WO2022223005A1 - 使用植物乳杆菌twk10来促进骨愈合的方法 - Google Patents

使用植物乳杆菌twk10来促进骨愈合的方法 Download PDF

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WO2022223005A1
WO2022223005A1 PCT/CN2022/088243 CN2022088243W WO2022223005A1 WO 2022223005 A1 WO2022223005 A1 WO 2022223005A1 CN 2022088243 W CN2022088243 W CN 2022088243W WO 2022223005 A1 WO2022223005 A1 WO 2022223005A1
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lactobacillus plantarum
composition
bone
individual
twk10
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PCT/CN2022/088243
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English (en)
French (fr)
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黄毓慈
周玄
吴俊贤
李珈嘉
许涵茵
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生合生物科技股份有限公司
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Priority claimed from TW108122113A external-priority patent/TWI774966B/zh
Application filed by 生合生物科技股份有限公司 filed Critical 生合生物科技股份有限公司
Priority to EP22791111.2A priority Critical patent/EP4327816A1/en
Priority to US17/915,436 priority patent/US20230139904A1/en
Publication of WO2022223005A1 publication Critical patent/WO2022223005A1/zh

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    • A—HUMAN NECESSITIES
    • A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
    • A61K—PREPARATIONS FOR MEDICAL, DENTAL OR TOILETRY PURPOSES
    • A61K35/00—Medicinal preparations containing materials or reaction products thereof with undetermined constitution
    • A61K35/66—Microorganisms or materials therefrom
    • A61K35/74—Bacteria
    • A61K35/741—Probiotics
    • A61K35/744—Lactic acid bacteria, e.g. enterococci, pediococci, lactococci, streptococci or leuconostocs
    • A61K35/747—Lactobacilli, e.g. L. acidophilus or L. brevis
    • A—HUMAN NECESSITIES
    • A23—FOODS OR FOODSTUFFS; TREATMENT THEREOF, NOT COVERED BY OTHER CLASSES
    • A23L—FOODS, FOODSTUFFS, OR NON-ALCOHOLIC BEVERAGES, NOT COVERED BY SUBCLASSES A21D OR A23B-A23J; THEIR PREPARATION OR TREATMENT, e.g. COOKING, MODIFICATION OF NUTRITIVE QUALITIES, PHYSICAL TREATMENT; PRESERVATION OF FOODS OR FOODSTUFFS, IN GENERAL
    • A23L33/00—Modifying nutritive qualities of foods; Dietetic products; Preparation or treatment thereof
    • A23L33/10—Modifying nutritive qualities of foods; Dietetic products; Preparation or treatment thereof using additives
    • A23L33/135—Bacteria or derivatives thereof, e.g. probiotics
    • A—HUMAN NECESSITIES
    • A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
    • A61P—SPECIFIC THERAPEUTIC ACTIVITY OF CHEMICAL COMPOUNDS OR MEDICINAL PREPARATIONS
    • A61P19/00—Drugs for skeletal disorders
    • A61P19/08—Drugs for skeletal disorders for bone diseases, e.g. rachitism, Paget's disease
    • A—HUMAN NECESSITIES
    • A23—FOODS OR FOODSTUFFS; TREATMENT THEREOF, NOT COVERED BY OTHER CLASSES
    • A23V—INDEXING SCHEME RELATING TO FOODS, FOODSTUFFS OR NON-ALCOHOLIC BEVERAGES AND LACTIC OR PROPIONIC ACID BACTERIA USED IN FOODSTUFFS OR FOOD PREPARATION
    • A23V2002/00—Food compositions, function of food ingredients or processes for food or foodstuffs
    • A—HUMAN NECESSITIES
    • A23—FOODS OR FOODSTUFFS; TREATMENT THEREOF, NOT COVERED BY OTHER CLASSES
    • A23V—INDEXING SCHEME RELATING TO FOODS, FOODSTUFFS OR NON-ALCOHOLIC BEVERAGES AND LACTIC OR PROPIONIC ACID BACTERIA USED IN FOODSTUFFS OR FOOD PREPARATION
    • A23V2400/00—Lactic or propionic acid bacteria
    • A23V2400/11—Lactobacillus
    • A23V2400/169—Plantarum
    • A—HUMAN NECESSITIES
    • A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
    • A61K—PREPARATIONS FOR MEDICAL, DENTAL OR TOILETRY PURPOSES
    • A61K35/00—Medicinal preparations containing materials or reaction products thereof with undetermined constitution
    • A61K2035/11—Medicinal preparations comprising living procariotic cells
    • A61K2035/115—Probiotics

Definitions

  • the present invention relates to the use of Lactobacillus plantarum TWK10 (CGMCC NO. 13008) to promote bone healing in an individual.
  • CGMCC NO. 13008 Lactobacillus plantarum TWK10
  • Taiwan TW I583388 B revealed that Lactobacillus plantarum BCRC 910734 can be used to increase exercise performance and reduce fatigue.
  • 6-week-old mice were found to have stronger forelimb grip strength and longer swimming times after taking Lactobacillus plantarum BCRC 910734 for 6 weeks, and Blood lactate and ammonia concentrations also decreased significantly after exercise.
  • Lactobacillus plantarum TWK10 (CGMCC NO.13008) can achieve bone healing by promoting the activation and generation of osteoblasts (osteoblast) and bone maturation (bone maturation). bone healing).
  • the present invention provides a method for promoting bone healing in an individual, comprising administering to an individual in need thereof a bacterium comprising Lactobacillus plantarum TWK10 (CGMCC NO. 13008). combination.
  • the present invention provides the use of Lactobacillus plantarum TWK10 (CGMCC NO. 13008) for the preparation of a composition for promoting bone healing in a subject.
  • CGMCC NO. 13008 Lactobacillus plantarum TWK10
  • the present invention provides a composition for promoting bone healing in a subject comprising Lactobacillus plantarum TWK10 (CGMCC NO. 13008).
  • the present invention provides a supply of Lactobacillus plantarum TWK10 (CGMCC NO. 13008) for promoting bone healing in an individual.
  • CGMCC NO. 13008 Lactobacillus plantarum TWK10
  • Figure 1 and Figure 2 respectively show the bone volume fraction (BV/TV) (%) and bone mineral density (BMD) (g/mm 3 of the femurs of the rats in the control group 1 and the experimental group 1 in Example 1) ), where "*" means: when compared with the control group 1, p ⁇ 0.05;
  • Fig. 3 shows the results observed by hematoxylin-eosin staining of the femoral tissue sections of the rats in each group of Example 1, wherein the red arrows indicate osteoblasts;
  • Figure 4 shows the ratio of activated osteoblasts (%) measured by the femoral tissue sections of the rats in each group of Example 1, wherein “*” and “***” respectively indicate: when compared with the corresponding control group , p ⁇ 0.05 and p ⁇ 0.001;
  • Figure 5 shows the area percentage (%) of lamellar bone measured by the femoral tissue sections of the rats in each group of Example 1, where "**” and "***” respectively represent: when compared with the corresponding control group p ⁇ 0.01 vs. p ⁇ 0.001; and
  • Figure 6 shows the percentage (%) of SATB2 expression measured by the femoral tissue sections of the rats in each group of Example 1, wherein “**” and “***” respectively indicate: when compared with the corresponding control group, p ⁇ 0.01 and p ⁇ 0.001.
  • the present invention provides a composition for promoting bone healing in an individual comprising Lactobacillus plantarum TWK10 (CGMCC NO. 13008).
  • the present invention also provides the use of Lactobacillus plantarum TWK10 (CGMCC NO. 13008) for the preparation of a composition for promoting bone healing in an individual.
  • the present invention further provides a method for promoting bone healing in an individual comprising administering to an individual in need thereof a composition comprising Lactobacillus plantarum TWK10.
  • administering and “administration” are used interchangeably and mean introducing, providing or providing to an individual by any suitable route. Delivering a predetermined active ingredient to perform its intended utility.
  • the present invention provides Lactobacillus plantarum TWK10 supply for promoting bone healing in an individual.
  • Lactobacillus plantarum TWK10 can be live or dead, concentrated or non-concentrated, liquid, paste, semi-solid ), or solid [eg, pellet, granule, or powder], and may be heat-inactivated, frozen, dried dried, or freeze-dried [e.g., in freeze-dried form or spray/fluid bed dried form].
  • Lactobacillus plantarum TWK10 exists in the form of freeze-dried powder.
  • bone healing and the term “bone repair” are used interchangeably.
  • the individual may be an individual with a fracture, and the fracture may be caused by at least one of the following: trauma; congenital bone healing disorders, eg, bone osteogenesis imperfecta; cancer, e.g., bone metastasis; and surgery, e.g., osteotomies [such as bone lengthening and limb lengthening procedures)] and tumor resection surgery (such as the resection of primary bone tumors and the resection of secondary bone cancers (also known as bone metastases)].
  • trauma congenital bone healing disorders, eg, bone osteogenesis imperfecta
  • cancer e.g., bone metastasis
  • surgery e.g., osteotomies [such as bone lengthening and limb lengthening procedures)]
  • tumor resection surgery such as the resection of primary bone tumors and the resection of secondary bone cancers (also known as bone metastases)].
  • the fractures may include, but are not limited to: simple fractures, open fractures, transverse fractures, fissure fractures (also known as longitudinal fractures), Oblique fracture (oblique fracture), spiral fracture (spiral fracture), compression fracture (compression fracture), segmental fracture (segmental fracture), comminuted fracture (communicated fracture), avulsion fracture (avulsion fracture), Complete fracture (greenstick fracture) and dislocation fracture (dislocation fracture).
  • the individual may be an individual with osteonecrosis, eg, osteonecrosis complicated by fractures, and ischemic osteonecrosis caused by osteomyelitis.
  • osteonecrosis eg, osteonecrosis complicated by fractures, and ischemic osteonecrosis caused by osteomyelitis.
  • the individual may be an individual undergoing a surgical intervention of bones, which may include, but is not limited to, bone replacement, such as a hip joint and Replacement of the knee joint; and bone implantation, eg, tooth implantation.
  • bones which may include, but is not limited to, bone replacement, such as a hip joint and Replacement of the knee joint; and bone implantation, eg, tooth implantation.
  • the individual is an individual without osteoporosis.
  • subject means any mammal of interest, such as humans, monkeys, cows, sheep, horses, Pigs, goats, dogs, cats, mice and rats.
  • the composition can be formulated into a food product using standard techniques well known to those of ordinary skill in the art.
  • the composition can be added directly to an edible material, or it can be used to prepare an intermediate composition suitable for subsequent addition to an edible material [eg, a food food additive or premix].
  • the term "food product” means any article or substance that can be ingested into the body of an individual.
  • examples of such food products may include, but are not limited to: fluid milk products such as milk and concentrated milk; fermented milk such as yogurt, sour milk and frozen yogurt; milk powder; ice cream; cheese (cream cheeses); dry cheese; soybean milk; fermented soybean milk); vegetable-fruit juices; fruit juices; sports drinks; jelly; cookies; energy bars; health foods; animals animal feeds; and dietary supplements.
  • the composition may also be prepared in the form of a pharmaceutical composition.
  • the pharmaceutical composition may be in a dosage form suitable for oral administration, parenteral administration or topical administration.
  • the pharmaceutical composition may further comprise a pharmaceutically acceptable carrier that is widely used in pharmaceutical manufacturing techniques.
  • the pharmaceutically acceptable carrier may contain one or more agents selected from the group consisting of: a solvent, a buffer, an emulsifier, a suspending agent, a disintegrating agent ( decomposer), disintegrating agent, dispersing agent, binding agent, excipient, stabilizing agent, chelating agent, diluent ), gelling agents, preservatives, wetting agents, lubricants, absorption delaying agents, liposomes and the like.
  • a solvent a buffer, an emulsifier, a suspending agent, a disintegrating agent ( decomposer), disintegrating agent, dispersing agent, binding agent, excipient, stabilizing agent, chelating agent, diluent ), gelling agents, preservatives, wetting agents, lubricants, absorption delaying agents, liposomes and the like.
  • the pharmaceutical composition can be manufactured into a dosage form suitable for oral administration using techniques well known to those skilled in the art, including, but not limited to: sterile powder, tablet, tablet troche, lozenge, pellet, capsule, dispersible powder or granule, solution, suspension, drop, emulsion (emulsion), syrup (syrup), elixir (elixir), thick syrup (slurry) and the like.
  • the pharmaceutical composition can be manufactured into a dosage form suitable for parenteral administration using techniques well known to those skilled in the art [including injections, e.g., sterile aqueous solutions]. solution or dispersion] and administered by a route selected from the group consisting of: intraperitoneal injection, intrapleural injection, intramuscular injection injection), intravenous injection, intraarterial injection, intraarticular injection, intrasynovial injection, intrathecal injection, intracranial injection injection), intraepidermal injection, subcutaneous injection, intradermal injection, intralesional injection, and sublingual administration.
  • a route selected from the group consisting of: intraperitoneal injection, intrapleural injection, intramuscular injection injection), intravenous injection, intraarterial injection, intraarticular injection, intrasynovial injection, intrathecal injection, intracranial injection injection), intraepidermal injection, subcutaneous injection, intradermal injection, intralesional injection, and sublingual administration.
  • the pharmaceutical composition can also be manufactured into an external preparation suitable for topical application to the skin using techniques well known to those skilled in the art, including, but not limited to: emulsions ), gel, ointment, cream, patch, liniment, powder, aerosol, spray, lotion ), serum, paste, foam, drop, suspension, salve and bandage.
  • Lactobacillus plantarum TWK10 will vary depending on the following factors: the severity of the disease to be treated, the route of administration, and the age, physical condition and response of the individual to be treated.
  • Lactobacillus plantarum TWK10 can be administered parenterally, orally or topically in a single dose or in divided doses.
  • Lactobacillus plantarum CGMCC NO.13008 1. Lactobacillus plantarum CGMCC NO.13008:
  • Lactobacillus plantarum CGMCC NO. 13008 also known as Lactobacillus plantarum TWK10 or Lactobacillus plantarum TWK10 used in the following examples has been approved by the Budapest Treaty on September 13, 2016 in accordance with the provisions of the Budapest Treaty. It has been deposited in the China General Microbiological Culture Collection Center (CGMCC), and is available to the public.
  • CGMCC China General Microbiological Culture Collection Center
  • Lactobacillus plantarum TWK10 was also deposited in the Biological Resources Conservation and Research Center of the Food Industry Research and Development Institute, FIRDI, Taiwan, China under the deposit number BCRC 910734 on June 30, 2016. Bioresource Collection and Research Center, BCRC) (No. 331, Food Road, Hsinchu City, Taiwan, China), and has been publicly available.
  • Wistar rats (6 weeks old, weighing approximately 200 to 260 g) used in the following examples were purchased from BioLasco Taiwan Co., Ltd. All experimental animals were housed in an individually air-conditioned animal room with 12 hours of light and dark, room temperature maintained at 21 ⁇ 2°C, and relative humidity maintained at 55 ⁇ 10%, and water and feed were adequately supplied. All experimental procedures related to laboratory animals are approved by the Institutional Animal Care and Use Committee (IACUC) of Fu Jen Democratic University (Fu Jen Democratic University), and are based on the National Institutes of Health. , NIH) of the 2011 8th edition of the Laboratory Animal Feeding Management and Use Specifications (Guide for the Care and Use of Laboratory Animals, 8th edition, 2011).
  • IACUC Institutional Animal Care and Use Committee
  • the bioceramic implants used for bone implantation in the following examples were provided by the laboratory of Professor Huang Yuci of the School of Medicine (Fu Jen Democratic University).
  • the bioceramic implant is a kind of silica powder (Nippon Light Metal Holdings Company, Ltd.) mixed with silica gel (Nissan Chemical America Corporation)
  • silica-based bioceramic scaffold which was fabricated with reference to Taiwan TW I421062 B and using 3D printing technology.
  • Example 1 Evaluation of the efficacy of Lactobacillus plantarum TWK10 in promoting bone healing
  • Lactobacillus plantarum TWK10 was inoculated into MRS medium and cultured at 37°C for 16-18 hours. After that, the resulting culture was centrifuged at 5,000 rpm for 10 minutes at 4°C, then the supernatant was removed, and the pellets were washed with an appropriate amount of physiological saline. After the bacterial count was carried out on the plate counting medium, freeze-drying was performed, and then an appropriate amount of maltodextrin was used for mixing to obtain a bacterial concentration of 10 10 CFU/g of Lactobacillus plantarum TWK10. Freeze-dried powder.
  • the rats of each experimental group were respectively administered with the freeze-dried bacterial powder of Lactobacillus plantarum TWK10 obtained in item A above (the dose is 10 9 CFU) /kg, in ddH 2 O), and the rats in each control group were administered with equal weight of maltodextrin (in ddH 2 O), wherein the rats in control group 1 and experimental group 1 were administered weekly
  • the rats in the control group 2 and the experimental group 2 were dosed twice a week for a total of 13 weeks.
  • the rats of each group were subjected to bone destruction and bone transplantation as follows: First, the rats of each group were anesthetized by inhalation of isoflurane. Next, the legs and abdomens of the rats in each group were shaving, followed by incision of the skin layer using sterile surgical scissors to form an incision, followed by a sterile scalpel The muscle beneath the skin layer was incised so that the femur beneath the muscle was exposed. After that, the femur was bone-destructed with a drill, so that a bore hole with a diameter of 2 mm was formed in the femur. Finally, a bioceramic implant is placed in the bore and sutured.
  • rats in control group 1 and experimental group 1 were sacrificed by CO 2 , and then the femurs were removed and used for experiments in items C to E below.
  • the rats of the control group 2 and the experimental group 2 were sacrificed by CO 2 as well, and then the femurs were taken out and used for the experiments in the following items D to E.
  • Rat femurs were photographed using a micro-computed tomography (micro-CT) system (SKYSCAN 1176, Bruker Corporation) at a magnification of 400 times, and then Five areas were randomly selected from the periphery of the bioceramic implant to measure bone volume (BV), tissue volume (TV), and bone mass density (BMD) (g /mm 3 ).
  • micro-CT micro-computed tomography
  • BMD bone mass density
  • Bone volume fraction (BV/TV) (%) was calculated by substituting the measured bone volume and tissue volume into the following formula (1):
  • A bone volume fraction (BV/TV) (%)
  • Rat femurs were fixation overnight using 10% formaldehyde solution followed by decalcification using 3% hydrochloric acid for 5 hours. Next, the fixed and decalcified femur was embedded in paraffin, and then sliced, thereby obtaining tissue sections having a thickness of 4 ⁇ m.
  • tissue section was stained by using hematoxylin-eosin and according to techniques well known and customary to those skilled in the art. Stained tissue sections were observed and photographed at a magnification of 400 times using a light microscope (Olympus BX51, Japan), and then activated osteoblasts and non-activated osteoblasts were counted, respectively. The number of cells (inactive osteoblast). The active osteoblast ratio (%) was calculated by substituting the obtained numbers of activated osteoblasts and non-activated osteoblasts into the following formula (2):
  • lamellar bone formation (lamellar bone formation) was observed and photographed under polarized light view, and analyzed using ImageJ Imaging Software (version 1.48). The area of lamellar bone surrounding the implantation of the bioceramic implant and the total tissue area, where the formation of lamellar bone represents the occurrence of bone maturation.
  • the lamellar bone area percentage (%) is calculated by substituting the measured lamellar bone area and total tissue area into the following formula (3):
  • G area percentage of layered bone (%)
  • tissue section obtained in item D above, and then use anti-special AT-rich sequence-binding protein 2 (SATB2) antibody (label Abcam, Cat. No. ab92446) as a primary antibody, and use the Rabbit specific HRP/DAB(ABC) Detection IHC Kit (the brand is Abcam, the product number is ab64261) and according to this Immunohistochemical staining is performed by techniques well known and routine to those skilled in the art. Stained tissue sections were observed and photographed using a light microscope (Olympus BX51, Japan) at a magnification of 400 times, followed by analysis using ImageJ imaging software (version 1.48), whereby the location of the tissue was measured.
  • SATB2 anti-special AT-rich sequence-binding protein 2
  • the bioceramic implant was implanted around the tissue area with SATB2 expression and the total tissue area, wherein the SATB2 expression represented the situation of osteoblastogenesis.
  • SATB2 expression percentage (%) was calculated by substituting the measured tissue area with SATB2 expression and the total tissue area into the following formula (4):
  • Figure 1 and Figure 2 respectively show the measured BV/TV and BMD of the femurs of the rats in the control group 1 and the experimental group 1 at the end of the 5th week after the start of administration. It can be seen from Figure 1 and Figure 2 that compared with the control group 1, the BV/TV and BMD measured in the experimental group 1 both increased significantly.
  • Fig. 3 shows the results observed by hematoxylin-eosin staining of femoral tissue sections of rats in each group at the end of the 5th week or the 13th week after the start of administration. It can be seen from Figure 3 that compared with control group 1, the femoral tissue of experimental group 1 obviously has more osteoblasts. Compared with control group 2, thicker bone formation could be observed in the femoral tissue of experimental group 2.
  • Fig. 4 shows the ratio (%) of activated osteoblasts measured in femoral tissue sections of rats in each group at the end of the 5th week or the 13th week after the start of administration. It can be seen from Figure 4 that the ratios of activated osteoblasts measured in each experimental group were significantly higher than those in the corresponding control group.
  • Figure 5 shows the area percentage (%) of lamellar bone measured by the femoral tissue sections of the rats in each group at the end of the 5th week or the 13th week after the start of administration. It can be seen from Fig. 5 that the area percentage of lamellar bone measured in each experimental group is significantly higher than that of the corresponding control group.
  • Fig. 6 shows the percentage (%) of SATB2 expression measured in femoral tissue sections of rats in each group at the end of the 5th week or the 13th week after the start of administration. It can be seen from Figure 6 that the percentage of SATB2 expression measured in each experimental group was significantly higher than that in the corresponding control group.
  • Lactobacillus plantarum TWK10 can effectively promote the activation and generation of osteoblasts and bone maturation whether it is administered for 5 weeks or 13 weeks. Therefore, the applicant believes that Lactobacillus plantarum TWK10 has a high potential for promoting bone healing.
  • Classification name Lactobacillus plantarum TWK10

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Abstract

提供了以植物乳杆菌TWK10(CGMCC NO.13008)用来促进骨愈合的方法。

Description

使用植物乳杆菌TWK10来促进骨愈合的方法 技术领域
本发明是有关于使用植物乳杆菌TWK10(CGMCC NO.13008)来促进一个体中的骨愈合。
背景技术
当骨骼受到外力而损伤[例如裂纹骨折(fissure fracture)(又称为骨裂)]时，主要是通过患者自身的骨愈合(bone healing)的能力来进行愈合，但当损伤程度较为严重而造成骨缺损(bone defects)[例如节断性骨折(segmental fracture)]时，则需进一步进行骨移植(bone implantation)来帮助骨愈合。而为了将植入物(implant)植入骨骼中，有时还需要先对患者进行骨破坏(bone destruction)。然而，骨愈合的能力会随着骨骼损伤的程度以及年龄的增加而相对地降低。因此，本领域的相关研究人员皆致力于发展出可以有效地促进骨愈合的手段。
中国台湾TW I583388 B揭示植物乳杆菌(Lactobacillus plantarum)BCRC 910734可被用来增加运动表达(exercise performance)及降低疲劳(fatigue)。在此件专利案的实施例中，6周大的小鼠在服用植物乳杆菌BCRC 910734历时6周后，被发现具有较强的前肢抓力(forelimb grip strength)以及较长的游泳时间，并且在运动后血液中乳酸与氨的浓度也有显著的降低。
发明内容
于本发明中，申请人意外地发现植物乳杆菌(Lactobacillus plantarum)TWK10(CGMCC NO.13008)能够通过促进成骨细胞(osteoblast)的活化与生成以及骨成熟(bone maturation)来达致骨愈合(bone healing)的效用。
于是，在第一个方面，本发明提供一种用来促进一个体中的骨愈合的方法，其包括对一有此需要的个体投予一包含有植物乳杆菌TWK10(CGMCC NO.13008)的组合物。
在第二个方面，本发明提供一种植物乳杆菌TWK10(CGMCC  NO.13008)供应用于制备一用来促进一个体中的骨愈合的组合物的用途。
在第三个方面，本发明提供一种用来促进一个体中的骨愈合的组合物，其包含有植物乳杆菌TWK10(CGMCC NO.13008)。
在第四个方面，本发明提供一种植物乳杆菌TWK10(CGMCC NO.13008)供应用于促进一个体中的骨愈合。
附图说明
图1与图2分别显示实施例1的对照组1与实验组1的大鼠股骨所测得的骨体积分率(BV/TV)(％)与骨质密度(BMD)(g/mm 3)，其中“*”表示：当与对照组1作比较，p＜0.05；
图3显示实施例1的各组大鼠的股骨组织切片通过苏木精-伊红染色而被观察到的结果，其中红色箭头指示代表成骨细胞；
图4显示实施例1的各组大鼠的股骨组织切片所测得的活化的成骨细胞比率(％)，其中“*”与“***”分别表示：当与对应的对照组作比较，p＜0.05与p＜0.001；
图5显示实施例1的各组大鼠的股骨组织切片所测得的层性骨的面积百分比(％)，其中“**”与“***”分别表示：当与对应的对照组作比较，p＜0.01与p＜0.001；以及
图6显示实施例1的各组大鼠的股骨组织切片所测得的SATB2表达百分比(％)，其中“**”与“***”分别表示：当与对应的对照组作比较，p＜0.01与p＜0.001。
具体实施方式
为了本说明书的目的，将被清楚地了解的是：文字“包含有(comprising)”意指“包含但不限于”，以及文字“包括(comprises)”具有一对应的意义。
除非另外有所定义，在本文中所使用的所有技术性与科学术语具有本领域技术人员所共同了解的意义。本领域技术人员会认知到许多与那些被描述于本文中者相似或等效的方法和材料，它们可被用于实施本发明。当然，本发明决不受到所描述的方法和材料的限制。
本发明提供一种用来促进一个体中的骨愈合(bone healing)的组 合物，其包含有植物乳杆菌TWK10(CGMCC NO.13008)。
本发明也提供一种植物乳杆菌TWK10(CGMCC NO.13008)供应用于制备一用来促进一个体中的骨愈合的组合物的用途。本发明又提供一种用来促进一个体中的骨愈合的方法，其包括对一有此需要的个体投予(administering)一包含有植物乳杆菌TWK10的组合物。
如本文中所使用的，术语“投予(administering)”以及“投药(administration)”可被交换地使用，并且意指通过任何合适的途径来对一个体导入(introducing)、提供(providing)或递送(delivering)一预定的活性成分以执行其预期的效用。
此外，本发明提供植物乳杆菌TWK10供应用于促进一个体中的骨愈合。
依据本发明，植物乳杆菌TWK10可以是活菌或死菌、经浓缩的(concentrated)或未经浓缩的(non-concentrated)、液态(liquid)、糊状(paste)、半固态(semi-solid)，或固态(solid)[例如，丸(pellet)、细颗粒(granule)或粉末(powder)]，并且可以是经热去活的(heat-inactivated)、经冷冻的(frozen)、经干燥的(dried)，或经冷冻干燥的(freeze-dried)[例如，可呈冷冻干燥形式或喷雾/流化床干燥(spray/fluid bed dried)形式]。在本发明的一个较佳具体例中，植物乳杆菌TWK10是呈冷冻干燥粉末的形式存在。
如本文中所使用的，术语“骨愈合(bone healing)”与术语“骨修补(bone repair)”可被交替地使用。
依据本发明，该个体可为一带有骨折(fracture)的个体，而该骨折可由下列中的至少一者所造成：外伤(trauma)；先天性骨愈合疾病(congenital bone healing disorders)，例如，骨生成不全症(osteogenesis imperfecta)；癌症(cancer)，例如，骨转移(bone metastasis)；以及手术(surgery)，例如，截骨术(osteotomies)[诸如骨延长与肢体延长手术(bone lengthening and limb lengthening procedures)]以及肿瘤切除术(tumor resection surgery)[诸如原发性骨肿瘤(primary bone tumor)的切除以及次发性骨癌(secondary bone cancer)(又称为骨转移)的切除]。
依据本发明，该骨折可包括，但不限于：简单骨折(simple fracture)、开放性骨折(open fracture)、横向骨折(transverse fracture)、 裂纹骨折(fissure fracture)(又被称为纵向骨折)、斜向骨折(oblique fracture)、螺旋骨折(spiral fracture)、压迫性骨折(compression fracture)、节断性骨折(segmental fracture)、粉碎性骨折(communicated fracture)、撕除性骨折(avulsion fracture)、不完全骨折(greenstick fracture)以及脱臼骨折(dislocation fracture)。
另择地，该个体也可为一带有骨坏死(osteonecrosis)的个体，例如，骨折并发的骨坏死，以及骨髓炎(osteomyelitis)导致的缺血性骨坏死(ischemic osteonecrosis)。
依据本发明，该个体可为一经历骨手术介入(surgical intervention of bones)的个体，而该骨手术介入可包括，但不限于：骨置换(bone replacement)，例如，髋关节(hip joint)与膝关节(knee joint)的置换；以及骨移植(bone implantation)，例如，植牙(tooth implantation)。
较佳地，该个体是一不带有骨质疏松(osteoporosis)的个体。
如本文中所使用的，术语“个体(subject)”意指任何感兴趣的哺乳类动物，诸如人(humans)、猴子(monkeys)、牛(cows)、绵羊(sheep)、马(horses)、猪(pigs)、山羊(goats)、狗(dogs)、猫(cats)、小鼠(mice)以及大鼠(rats)。
依据本发明，该组合物可使用具有本领域普通技术人员所熟知的标准技术而被配制成一食品产品(food product)。例如，该组合物可直接地被添加至一可食性材料(edible material)内，或者它可被用于制备一适用于随后添加至可食性材料内的中间组合物(intermediate composition)[例如，食品添加物(food additive)或预混料(premix)]。
如本文中所使用的，术语“食品产品(food product)”意指可以被一个体摄入至其体内的任何物品(article)或物质(substance)。依据本发明，该食物产品的实例可包括，但不限于：流体乳品(fluid milk products)，例如牛奶(milk)以及浓缩牛奶(concentrated milk)；发酵乳品(fermented milk)，例如酸奶(yogurt)、酸乳(sour milk)以及冷冻优格(frozen yogurt)；奶粉(milk powder)；冰淇淋(ice cream)；奶酪(cream cheeses)；干酪(dry cheeses)；豆奶(soybean milk)；发酵豆奶(fermented soybean milk)；蔬果汁(vegetable-fruit juices)；果汁(fruit juices)；运动饮料(sports drinks)；果冻(jelly)；饼干(cookies)；能量棒(energy bars)； 健康食品(health foods)；动物饲料(animal feeds)；以及膳食补充品(dietary supplements)。
依据本发明，该组合物也可以一药学组合物(pharmaceutical composition)的形式而被制得。
依据本发明，该药学组合物可呈一适合于口服投药(oral administration)、非经肠道投药(parenteral administration)或局部投药(topical administration)的剂型(dosage form)。
依据本发明，该药学组合物可进一步包含有一被广泛地使用于药物制造技术的药学上可接受的载剂(pharmaceutically acceptable carrier)。例如，该药学上可接受的载剂可包含一或多种选自于下列的试剂：溶剂(solvent)、缓冲液(buffer)、乳化剂(emulsifier)、悬浮剂(suspending agent)、分解剂(decomposer)、崩解剂(disintegrating agent)、分散剂(dispersing agent)、黏结剂(binding agent)、赋形剂(excipient)、稳定剂(stabilizing agent)、螯合剂(chelating agent)、稀释剂(diluent)、胶凝剂(gelling agent)、防腐剂(preservative)、润湿剂(wetting agent)、润滑剂(lubricant)、吸收延迟剂(absorption delaying agent)、脂质体(liposome)以及类似物。有关这些试剂的选用与数量是落在本领域技术人员的专业素养与例行技术范畴内。
依据本发明，该药学组合物可利用本领域技术人员所详知的技术而被制造成一适合于口服投药的剂型，这包括，但不限于：无菌的粉末、锭剂(tablet)、片剂(troche)、口含锭(lozenge)、丸剂(pellet)、胶囊(capsule)、分散性粉末(dispersible powder)或细颗粒(granule)、溶液、悬浮液(suspension)、滴剂(drop)、乳剂(emulsion)、糖浆(syrup)、酏剂(elixir)、浓浆(slurry)以及类似物。
依据本发明，该药学组合物可利用本领域技术人员所详知的技术而被制造成一适合于非经肠道投药的剂型[包括注射品(injection)，例如，无菌的水性溶液(sterile aqueous solution)或分散液(dispersion)]，且以一选自于由下列所构成的群组中的途径来投药：腹膜内注射(intraperitoneal injection)、胸膜内注射(intrapleural injection)、肌肉内注射(intramuscular injection)、静脉内注射(intravenous injection)、动脉内注射(intraarterial injection)、关节内注射(intraarticular injection)、 滑液内注射(intrasynovial injection)、椎管内注射(intrathecal injection)、颅内注射(intracranial injection)、表皮内注射(intraepidermal injection)、皮下注射(subcutaneous injection)、皮内注射(intradermal injection)、病灶内注射(intralesional injection)、以及舌下投药(sublingual administration)。
依据本发明，该药学组合物也可利用本领域技术人员所详知的技术而被制造成一适合于局部地施用于皮肤上的外部制剂(external preparation)，这包括，但不限于：乳剂(emulsion)、凝胶(gel)、软膏(ointment)、乳霜(cream)、贴片(patch)、擦剂(liniment)、粉末(powder)、气溶胶(aerosol)、喷雾(spray)、乳液(lotion)、乳浆(serum)、糊剂(paste)、泡沫(foam)、滴剂(drop)、悬浮液(suspension)、油膏(salve)以及绷带(bandage)。
依据本发明，植物乳杆菌TWK10的投药剂量与投药次数会视下列因素而变化：要被治疗的疾病的严重性，投药途径，以及要被治疗的个体的年龄、身体状况与反应。一般而言，植物乳杆菌TWK10可呈单一剂量或是分成多个剂量的形式而被非经肠地道、口服地或局部地投药。
本发明将就下面的实施例来做进一步说明，但应了解的是，该等实施例只是供例示说明用，而不应被解释为本发明的实施上的限制。
＜实施例＞
一般实验材料：
1.植物乳杆菌(Lactobacillus plantarum)CGMCC NO.13008：
在下面实施例中所使用的植物乳杆菌CGMCC NO.13008(又被称为植物乳杆菌TWK10或胚芽乳杆菌TWK10)已有依据布达佩斯条约(the Budapest Treaty)的规定，于公元2016年9月13日被保藏于中国微生物菌种保藏管理委员会普通微生物中心(China General Microbiological Culture Collection Center,CGMCC)，并且已公众可获得。
另外，该植物乳杆菌TWK10也有以保藏编号BCRC 910734于公元2016年6月30日被保藏于中国台湾的食品工业发展研究所(Food Industry Research and Development Institute,FIRDI)的生物资源保存 及研究中心(Bioresource Collection and Research Center,BCRC)(300新竹市食品路331号，中国台湾)，并且已公众可获得。
2.实验动物：
在下面实施例中所使用的Wistar大鼠(Wistar rats)(6周大，体重约为200至260g)是购自于乐斯科生物科技股份有限公司(BioLasco Taiwan Co.,Ltd)。所有的实验动物被饲养于一个光照与黑暗各为12小时、室温维持在21±2℃以及相对湿度维持在55±10％的独立空调的动物房内，而且水分与饲料被充分地供给。有关实验动物的一切实验程序是由辅仁大学(Fu Jen Catholic University)的实验动物照护及使用委员会(Institutional Animal Care and Use Committee,IACUC)所认可，并依据美国国家卫生研究院(National Institutes of Health,NIH)的2011年第8版的实验动物饲养管理及使用规范(Guide for the Care and Use of Laboratory Animals,8th edition,2011)来进行。
3.生物陶瓷植入物(bioceramic implant)：
下面实施例中被使用来进行骨移植(bone implantation)的生物陶瓷植入物是由辅仁大学医学系(School of Medicine,Fu Jen Catholic University)的黄毓慈教授的研究室所提供。该生物陶瓷植入物是一种由二氧化硅粉末(silica powder)(Nippon Light Metal Holdings Company,Ltd.)与硅胶(silica gel)(Nissan Chemical America Corporation)所混合而成之以二氧化硅为基础的生物陶瓷支架(silica‐based bioceramic scaffold)，其是参考中国台湾TW I421062 B并使用3D打印技术(3D printing technology)而被制得。
一般实验方法：
1.统计学分析(statistical analysis)：
在下面的实施例中，各组的实验数据是以“平均值(mean)±标准误差(standard error,SE)”来表示。所有的数据是通过曼-惠特尼U检定(Mann-Whitney U test)来作分析，俾以评估各组间的差异性。若所得到的统计分析结果是p<0.05，这表示有统计学显著性(statistical significance)。
实施例1.植物乳杆菌TWK10在促进骨愈合(bone healing)上的效用评估
实验方法：
A.植物乳杆菌TWK10的冷冻干燥菌粉的制备：
首先，将植物乳杆菌TWK10接种至MRS培养基中，并于37℃下进行培养历时16-18小时。之后，所得到的培养物在4℃下以5,000rpm进行离心历时10分钟，接着去除上清液，而沉淀物(pellets)以适量的生理食盐水予以洗涤。在以平板计数培养基来进行菌数计数后，进行冷冻干燥处理，继而使用适量的麦芽糊精(maltodextrin)来予以混合，借此而得到细菌浓度为10 10CFU/g的植物乳杆菌TWK10的冷冻干燥菌粉。
B.植物乳杆菌TWK10的投药以及骨移植：
首先，将Wistar大鼠随机地分成4组(每组n＝4)，其中包括2个对照组(也就是对照组1与对照组2)以及2个实验组(也就是实验组1与实验组2)。接着，通过口服胃管灌食法(oral gavage)的方式，对各个实验组的大鼠分别投药以依据上面第A项当中所得到的植物乳杆菌TWK10的冷冻干燥菌粉(剂量为10 9CFU/kg，配于ddH 2O中)，以及对各个对照组的大鼠分别投药以等重量的麦芽糊精(配于ddH 2O中)，其中对照组1与实验组1的大鼠每周被投药5次，投药时间总共历时5周，至于对照组2与实验组2的大鼠则是每周被投药2次，投药时间总共历时13周。
在开始投药后的第1周结束时，对各组大鼠进行如下所述的骨破坏以及骨移植：首先，各组大鼠是通过异氟醚(isoflurane)的吸入而被麻醉。接着，将各组大鼠的腿腹部(abdomen)进行剃毛(shaving)，继而使用无菌的手术剪刀(surgical scissor)将皮肤层切开以形成切口(incision)，然后使用无菌的解剖刀(scalpel)将位于该皮肤层下方的肌肉切开而使得在该肌肉下方的股骨(femur)被显露出来。之后，以钻孔器(drill)来对该股骨进行骨破坏，而使得该股骨形成有一直径为2mm的钻孔(bore hole)。最后，将一生物陶瓷植入物植入该钻孔中并进行缝合。
在开始投药后的第5周结束时，通过CO 2来牺牲对照组1与实验组1的大鼠，接着取出股骨并将其拿来进行下面第C至E项的实验。另外，在开始投药后的第13周结束时，同样通过CO 2来牺牲对照组 2与实验组2的大鼠，接着取出股骨并将其拿来进行下面第D至E项的实验。
C.骨质参数(bone quality parameters)的分析：
使用微型-计算机断层摄影(micro-CT)系统[micro-computed tomography(micro-CT)system](SKYSCAN 1176,Bruker Corporation)并在一为400倍的放大倍率下来对大鼠的股骨进行拍照，然后从该生物陶瓷植入物植入的周围随机地选出5个区域来测量骨体积(bone volume,BV)、组织体积(tissue volume,TV)以及骨质密度(bone mass density,BMD)(g/mm 3)。
骨体积分率(bone volume fraction)(BV/TV)(％)是通过将所测得的骨体积与组织体积代入下列公式(1)而被计算出：
公式(1)：A＝(B/C)×100
其中：A＝骨体积分率(BV/TV)(％)
B＝骨体积(mm 3)
C＝组织体积(mm 3)
之后，依照上面“一般实验方法”的第1项「统计学分析」当中所述的方法来分析所得到的实验数据。
D.组织病理学分析(histopathological analysis)：
使用10％甲醛(formaldehyde)溶液来对大鼠的股骨进行固定处理(fixation)至隔夜，继而使用3％盐酸(hydrochloric acid)来进行脱钙(decalcification)历时5小时。接着，将经固定且脱钙的股骨以石蜡(paraffin)予以包埋(embedding)，然后进行切片处理，借此而得到具有一厚度为4μm的组织切片(tissue sections)。
之后，一部分的组织切片通过使用苏木精-伊红(hematoxylin-eosin)并且依据本领域技术人员所详知且惯用的技术来进行染色。经染色的组织切片是通过使用一光学显微镜(Olympus BX51,Japan)并在一为400倍的放大倍率下来进行观察以及拍照，然后分别计数活化的成骨细胞(active osteoblast)以及未活化的成骨细胞(inactive osteoblast)的数目。活化的成骨细胞比率(active osteoblast ratio)(％)是通过将所得到的活化的成骨细胞数目以及未活化的成骨细胞数目代入下列公式(2)而被计算出：
公式(2)：D＝[E/(E+F)]×100
其中：D＝活化的成骨细胞比率(％)
E＝活化的成骨细胞数目(个)
F＝未活化的成骨细胞数目(个)
另外，在偏光视野(polarized light view)下进行层性骨形成(lamellar bone formation)的观察以及拍照，并使用ImageJ成像软件(ImageJ Imaging Software，版本1.48)来进行分析，借此而测得位于该生物陶瓷植入物植入的周围的层性骨面积以及总组织面积，其中层性骨的形成代表有骨成熟(bone maturation)的发生。层性骨的面积百分比(lamellar bone area percentage)(％)是通过将所测得的层性骨面积以及总组织面积代入下列公式(3)而被计算出：
公式(3)：G＝(H/I)×100
其中：G＝层性骨的面积百分比(％)
H＝层性骨面积(mm 2)
I＝总组织面积(mm 2)
之后，依照上面“一般实验方法”的第1项「统计学分析」当中所述的方法来分析所得到的实验数据。
E.免疫组织化学染色(immunohistochemistry stain)：
收取上面第D项当中所得到的另一部分的组织切片，接着使用抗-特殊的富含AT的序列结合蛋白2(Special AT-rich sequence-binding protein 2,SATB2)抗体(厂牌为Abcam，货号为ab92446)作为一次抗体，以及使用兔子专一性HRP/DAB(ABC)侦测IHC试剂盒[Rabbit specific HRP/DAB(ABC)Detection IHC Kit](厂牌为Abcam，货号为ab64261)并且依据本领域技术人员所详知且惯用的技术来进行免疫组织化学染色。经染色的组织切片是通过使用光学显微镜(Olympus BX51,Japan)并在一为400倍的放大倍率下来进行观察以及拍照，继而使用ImageJ成像软件(版本1.48)来进行分析，借此而测得位于该生物陶瓷植入物植入的周围之具有SATB2表达的组织面积以及总组织面积，其中SATB2的表达代表有成骨细胞生成(osteoblastogenesis)的情形。SATB2表达百分比(SATB2 expression percentage)(％)是通过将所测得之具有SATB2表达的组织面积以及总组织面积代入下列公式 (4)而被计算出：
公式(4)：J＝(K/L)×100
其中：J＝SATB2表达百分比(％)
K＝具有SATB2表达的组织面积(mm 2)
L＝总组织面积(mm 2)
之后，依照上面“一般实验方法”的第1项「统计学分析」当中所述的方法来分析所得到的实验数据。
结果：
A.骨质参数的分析：
图1与图2分别显示在开始投药后的第5周结束时，对照组1与实验组1的大鼠股骨所测得的BV/TV与BMD。从图1与图2可见，与对照组1相较下，实验组1所测得的BV/TV与BMD皆有显著的增加。
B.组织病理学分析：
图3显示在开始投药后的第5周或第13周结束时，各组大鼠的股骨组织切片通过苏木精-伊红染色而被观察到的结果。从图3可见，与对照组1相较下，实验组1的股骨组织明显具有较多的成骨细胞。而与对照组2相较下，实验组2的股骨组织中能够观察到有较厚的骨形成。
图4显示在开始投药后的第5周或第13周结束时，各组大鼠的股骨组织切片所测得的活化的成骨细胞比率(％)。从图4可见，各个实验组所测得的活化的成骨细胞比率皆是显著高于对应的对照组所具者。
图5显示在开始投药后的第5周或第13周结束时，各组大鼠的股骨组织切片所测得的层性骨的面积百分比(％)。从图5可见，各个实验组所测得的层性骨的面积百分比皆是显著高于对应的对照组所具者。
C.免疫组织化学染色：
图6显示在开始投药后的第5周或第13周结束时，各组大鼠的股骨组织切片所测得的SATB2表达百分比(％)。从图6可见，各个实验组所测得的SATB2表达百分比皆是显著高于对应的对照组所具者。
综合以上的实验结果可知：无论是历时5周或13周的投药，植物乳杆菌TWK10皆能够有效地促进成骨细胞的活化与生成以及骨成熟。因此，申请人认为：植物乳杆菌TWK10具有用于促进骨愈合的高潜力。
于本说明书中被引述的所有专利和文献以其整体被并入本案作为参考数据。若有所冲突时，本案详细说明(包含界定在内)将占上风。
虽然本发明已参考上述特定的具体例被描述，明显地在不背离本发明的范围和精神下可作出很多的修改和变化。因此意欲的是，本发明仅受如随文检附的权利要求书所示者的限制。
生物材料保藏信息说明
保藏编号：CGMCC No.13008
分类命名：植物乳杆菌Lactobacillus plantarum TWK10
保藏日期：2016年9月13日
保藏单位：中国微生物菌种保藏管理委员会普通微生物中心
保藏单位地址：北京市朝阳区北辰西路1号院3号

Claims (10)

  1. 一种用来促进一个体中的骨愈合的方法，其特征在于：该方法包括对一有此需要的个体投予一包含有植物乳杆菌TWK10的组合物，所述植物乳杆菌TWK10是以保藏编号CGMCC NO.13008被保藏于中国微生物菌种保藏管理委员会普通微生物中心。
  2. 根据权利要求1所述的方法，其特征在于：该组合物是一食品产品或一药学组合物。
  3. 根据权利要求2所述的方法，其特征在于：该药学组合物是呈一供口服投药、非经肠道投药或局部投药的剂型。
  4. 一种植物乳杆菌TWK10供应用于制备一用来促进一个体中的骨愈合的组合物的用途，所述植物乳杆菌TWK10是以保藏编号CGMCC NO.13008被保藏于中国微生物菌种保藏管理委员会普通微生物中心。
  5. 根据权利要求4所述的用途，其特征在于：该组合物是一食品产品或一药学组合物。
  6. 根据权利要求5所述的用途，其特征在于：该药学组合物是呈一供口服投药、非经肠道投药或局部投药的剂型。
  7. 一种用来促进一个体中的骨愈合的组合物，其特征在于：该组合物包含有植物乳杆菌TWK10，所述植物乳杆菌TWK10是以保藏编号CGMCC NO.13008被保藏于中国微生物菌种保藏管理委员会普通微生物中心。
  8. 根据权利要求7所述的组合物，其特征在于：该组合物是一食品产品或一药学组合物。
  9. 根据权利要求8所述的组合物，其特征在于：该药学组合物是呈一供口服投药、非经肠道投药或局部投药的剂型。
  10. 一种植物乳杆菌TWK10供应用于促进一个体中的骨愈合，所述植物乳杆菌TWK10是以保藏编号CGMCC NO.13008被保藏于中国微生物菌种保藏管理委员会普通微生物中心。
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Citations (6)

* Cited by examiner, † Cited by third party
Publication number Priority date Publication date Assignee Title
TWI421062B (zh) 2009-10-21 2014-01-01 Univ Lunghwa Sci & Technology 多孔性生醫陶瓷骨骼支架之成型方法及成型設備
TWI583388B (zh) 2016-08-24 2017-05-21 國立體育大學 胚芽乳酸桿菌用於製備改善運動表現及降低肌肉疲勞之益生菌組合物之用途
CN108624520A (zh) * 2017-03-16 2018-10-09 景岳生物科技股份有限公司 促进骨质再生的植物乳杆菌菌株gmnl-662及其组合物
US20190321420A1 (en) * 2018-04-24 2019-10-24 Grape King Bio Ltd. Method and composition for preventing, treating or relieving bone diseases
WO2020175869A2 (ko) * 2019-02-26 2020-09-03 전남대학교산학협력단 신규한 락토바실러스 사케이 cvl-001 균주 또는 이의 배양액을 포함하는 골 질환 또는 대사성 질환 개선, 예방 또는 치료용 조성물
CN111621449A (zh) * 2020-07-02 2020-09-04 重庆第二师范学院 一种益生菌及其在继发性骨质疏松中的应用

Family Cites Families (2)

* Cited by examiner, † Cited by third party
Publication number Priority date Publication date Assignee Title
TWI604052B (zh) * 2017-02-20 2017-11-01 景岳生物科技股份有限公司 促進骨質再生的植物乳桿菌菌株ｇｍｎｌ－６６２及其組合物
TWI774966B (zh) * 2019-06-25 2022-08-21 生合生物科技股份有限公司 胚芽乳桿菌bcrc 910734用於提升老年個體的肌力以及治療肌肉減少症的用途

Patent Citations (6)

* Cited by examiner, † Cited by third party
Publication number Priority date Publication date Assignee Title
TWI421062B (zh) 2009-10-21 2014-01-01 Univ Lunghwa Sci & Technology 多孔性生醫陶瓷骨骼支架之成型方法及成型設備
TWI583388B (zh) 2016-08-24 2017-05-21 國立體育大學 胚芽乳酸桿菌用於製備改善運動表現及降低肌肉疲勞之益生菌組合物之用途
CN108624520A (zh) * 2017-03-16 2018-10-09 景岳生物科技股份有限公司 促进骨质再生的植物乳杆菌菌株gmnl-662及其组合物
US20190321420A1 (en) * 2018-04-24 2019-10-24 Grape King Bio Ltd. Method and composition for preventing, treating or relieving bone diseases
WO2020175869A2 (ko) * 2019-02-26 2020-09-03 전남대학교산학협력단 신규한 락토바실러스 사케이 cvl-001 균주 또는 이의 배양액을 포함하는 골 질환 또는 대사성 질환 개선, 예방 또는 치료용 조성물
CN111621449A (zh) * 2020-07-02 2020-09-04 重庆第二师范学院 一种益生菌及其在继发性骨质疏松中的应用

Non-Patent Citations (1)

* Cited by examiner, † Cited by third party
Title
HUANG, WENJING ET AL.: "Effect of Lactobacillus Plantarum TWK10 on Improving Endurance Performance in Humans", CHINESE JOURNAL OF PHYSIOLOGY, vol. 61, no. 3, 30 June 2018 (2018-06-30), XP055770695, ISSN: 0304-4920, DOI: 10.4077/CJP.2018.BAH587 *

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