CN115429933A - Application of BMSCs captured based on bio-orthogonal click chemistry reaction in bone field - Google Patents
Application of BMSCs captured based on bio-orthogonal click chemistry reaction in bone field Download PDFInfo
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Abstract
The invention belongs to the field of medicine, particularly relates to the field of bones, and particularly relates to application of capturing BMSCs based on bio-orthogonal click chemistry reaction in the field of bones. First of all, a titanium implant is disclosed, by grafting N, an azide group 3 The bone marrow mesenchymal stem cells are combined with the mussel bionic polypeptide modified titanium material to obtain the titanium implant. The invention designs and discovers a novel titanium implant which is based on a biological orthogonal click chemical reactionThe obtained product can fully exert osteogenic differentiation of stem cells without interfering normal physiological functions of cells, has more stable properties, can be used for promoting osseointegration and bone regeneration and repair, and brings good news for patients with bone injury.
Description
Technical Field
The invention belongs to the field of medicine, particularly relates to the field of bones, and particularly relates to application of a biological orthogonal click chemistry reaction in the field of bones.
Background
Titanium-based materials are the most commonly used biomedical metal materials, but the titanium materials have biological inertness, and implanted into bone tissues often causes weak bone forming capability of a titanium-bone interface, tissue fibrosis, particularly loose bone tissues, and the titanium implants are easy to generate aseptic loosening and the like. How to modify the inert surface of titanium material into a bioactive surface, promoting the osseointegration of titanium implant is a hot spot of biomaterial research.
Mesenchymal Stem Cells (BMSCs) in Bone marrow are important seed cells participating in the interfacial osteointegration of the titanium implant, and the BMSCs can be recruited to the surface of the titanium implant and promote Bone differentiation, finally form continuous Bone tissues on the surface of the titanium implant, and improve the bonding force of the implant-Bone interface. Therefore, titanium implants need to rely on the capacity of BMSCs to promote bone differentiation to achieve good osteointegration. In addition, it can be used in the field of bone regeneration repair.
Bio-orthogonal click chemistry is a completely new area. Bioorthogonal reactions, originally proposed by the professor Bertozzi in the united states, refer to a class of chemical reactions that can be performed in living organisms without interfering with the normal physiochemical processes of the living organisms. The bioorthogonal functional group modification is a metabolic modification strategy with strong functions, sugar, lipid and amino acid analogues containing functional groups such as azide groups, alkyne and aldehyde groups are added into a cell or organism culture solution through cell metabolic engineering, the structural analogues replace natural molecules such as carbohydrate and lipid through the metabolism of an organism, and then the structural analogues and a marker carrying a reactive group undergo click chemical reaction to form stable covalent connection, and finally the chemical marking of the functional groups is realized. The cell metabolism engineering utilizes the natural metabolic process of living bodies or cells, has little influence on the biological activity of a series of intermediate products and final products of the cell metabolic process, and is an ideal non-destructive modification mode.
The mussel bionic polypeptide modified titanium material surface is a simple and efficient method, and the bionic polypeptide carrying bioactive macromolecules can be grafted to the titanium material surface by a one-step soaking method to form a bioactive surface, so that the osteointegration promotion and bone regeneration repair effects of bioactive molecules are exerted. However, this modification method, which relies on bioactive molecules, is susceptible to environmental influences, is volatile, degrades, etc., and the number of BMSCs is limited, whereas the number of stem cells recruited and entered the titanium-bone interface after the titanium material is implanted into bone tissue is much smaller. Therefore, it is highly desirable to invent a new titanium material that can be used in the bone field.
Disclosure of Invention
The invention designs a strategy for promoting the osseointegration and the bone regeneration and repair of a titanium implant based on bioorthogonal reaction, which is characterized in that a more stable bioorthogonal reaction chemical group which is not influenced by the environment and does not interfere the normal physiological function of cells replaces a bioactive molecule to realize the capture of mesenchymal stem cells and combine the mesenchymal stem cells with the surface of a material, so that the osteogenic differentiation of the stem cells is exerted, and the osseointegration and the bone regeneration and repair are promoted.
Specifically, the technical scheme of the invention is as follows:
the first aspect of the invention discloses a titanium implant grafted with N azide group 3 The bone marrow mesenchymal stem cells are combined with the mussel bionic polypeptide modified titanium material to obtain the titanium implant.
Preferably, the bone marrow mesenchymal stem cells are captured and bonded on the surface of the titanium material through the bio-orthogonal click chemistry reaction of the azide group and the DBCO group on the surface of the titanium material.
Preferably, the titanium implant is a titanium screw.
In a second aspect, the invention discloses a method for promoting osseointegration and/or bone regeneration and repair based on bio-orthogonal click chemistry reaction, wherein a titanium implant obtained by the bio-orthogonal click chemistry reaction is implanted into an animal body to promote the osseointegration and/or bone regeneration and repair.
Preferably, N is grafted with an azide group 3 The bone marrow mesenchymal stem cells are combined with the mussel bionic polypeptide modified titanium material to obtain the titanium implant.
Preferably, a plurality ofThe sugar treatment of the mesenchymal stem cells leads the surfaces of the mesenchymal stem cells to be grafted with azide groups N 3 。
More preferably, the concentration of the polysaccharide is 0 to 100. Mu. Mol/L.
In some embodiments of the invention, the concentration of the polysaccharide is 20-30. Mu. Mol/L.
Preferably, the DBCO-modified mussel biomimetic polypeptide is synthesized by Fmoc solid phase synthesis.
More preferably, the mussel biomimetic polypeptide is grafted to the surface of the titanium material by coordination of DOPA groups and TiO2 on the surface of titanium.
The third aspect of the invention discloses the application of the titanium implant in the bone field. Preferably, the use of the titanium implant described above in osseointegration and/or bone regenerative repair is disclosed.
Compared with the prior art, the invention at least has the following beneficial effects:
the novel titanium implant is designed and found based on bioorthogonal click chemical reaction, does not interfere normal physiological functions of cells, can fully exert osteogenic differentiation of stem cells, has more stable properties, can be used for promoting osseointegration and bone regeneration and repair, and brings good news to patients with bone injury.
Drawings
FIG. 1 is a graph showing the effect of polysaccharide on the expression of sternness markers in rat BMSCs cells. Detecting the expression results (A) and further quantitative analysis results (B) of CD90, CD29, CD34 and CD45 on the surfaces of BMSCs after the treatment of different concentrations of polysaccharides by flow cytometry. (n =3, "+" indicates a statistical difference, "ns" indicates no statistical difference between the groups)
FIG. 2 is a graph showing the effect of polysaccharide on rat BMSCsALP activity. ALP staining results (A) and ALP activity assays (B) 7 days after osteogenic induction of rat BMSCs. (n =3, "+" indicates statistical differences,. + -. P <0.05,. + -. P < 0.01)
FIG. 3 is a graph showing the effect of polysaccharides on calcium nodule formation in rat BMSCs. Alizarin red staining results (a) and alizarin red quantification analysis (B) after 14 days of osteogenic induction of rat BMSCs. (n =3, "+" indicates that there is a statistical difference, P <0.05, "+ -P <0.01," + -P < 0.001)
FIG. 4 is a graph showing the effect of polysaccharides on the expression of rat BMSCs osteogenic genes. Expression of osteogenesis associated genes Alp (A) and Opn (B) 7 days after osteogenic induction in rat BMSCs. (n =3, "+" indicates statistical differences,. + -. P <0.05,. + -. P <0.01,. + -. P < 0.0001)
FIG. 5 is a graph showing the effect of polysaccharides on cartilage differentiation in rat BMSCs. Toluidine blue staining results after 14 days of chondrogenic induction of rat BMSCs.
FIG. 6 is a graph showing the effect of polysaccharides on adipogenic differentiation of rat BMSCs. After 14 days of rat BMSCs adipogenic induction, oil red O staining results (A) and PCR detection of the expression of Adiponectin (B) and Ppar-gamma (C) genes related to adipogenesis. (n =3, "+" indicates that there is a statistical difference, P <0.05, "+" P < 0.0001)
FIG. 7 is a schematic diagram showing the color development of polysaccharide treated rat BMSCs after binding to fluorescent probes.
FIG. 8 shows the fluorescence quantification of polysaccharide treated rat BMSCs bound to fluorescent probes at different concentrations. Flow cytometry (A) measured the fluorescence intensity and quantitative results (B) of different concentrations of polysaccharide-treated rat BMSCs reacted with fluorescent probes. (n =3, "+" indicates that there is a statistical difference, P <0.05, "+" P < 0.0001)
FIG. 9 is mass spectrometry analysis of biomimetic polypeptide EI-MS designed and synthesized. The analysis showed that the synthesized polypeptide [ M-H ] -was 1825.0.
FIG. 10 shows the effect of biomimetic polypeptide modification on various elements on the surface of titanium material. XPS (a) and EDS (B) characterize the elemental composition of the surface chemistry of the titanium sheet, with an N element content of 4.95% for the control group and 8.49% for the experimental group, and an increase in the N/Ti atomic ratio from 0.75 to 2.46 (N = 3).
FIG. 11 shows the effect of biomimetic polypeptide modification on the roughness of the surface of a titanium material. And observing the surface morphology result of the titanium sheet by using an atomic force microscope.
FIG. 12 shows the effect of biomimetic polypeptide modification on hydrophilicity and hydrophobicity of the surface of a titanium material. Measuring the water contact angle of the common titanium sheet and the titanium sheet grafted by the polypeptide (A) and quantifying the result (B). (n =3, "+" indicates a statistical difference,. + -. P < 0.01)
FIG. 13 shows BMSC-N 3 Calcium nodule formation ability after binding to DOPA-DBCO on titanium surface. Osteogenic induction for 14 and 21 days, alizarin red staining (a) and alizarin red quantification (B). (n =3, "ns" indicates that there is no statistical difference)
Fig. 14 is an image evaluation of the titanium screw-bone interface. And (5) carrying out Micro CT scanning to quantitatively analyze the osseointegration condition around the screw. (n =5, "+" indicates statistical differences,. + -. P <0.05,. + -. P <0.01,. + -. P <0.001,. + -. P < 0.0001)
Fig. 15 is a histological evaluation of the titanium screw-bone interface. Hard tissue section titanium screw-bone interface morphology and quantitative analysis results. (n =3, "+" indicates having statistical differences,. + -. P <0.01,. + -. P <0.001,. + -. P < 0.0001)
Fig. 16 shows the results of the titanium screw pullout resistance test. In particular to the analysis result of the load displacement curve and the maximum pull-out resistance force of each group of titanium screws. (n =3, "+" indicates that there is a statistical difference, P <0.05, "+" P < 0.0001)
Detailed Description
The technical solutions of the present invention are described in detail below with reference to the drawings and embodiments, but the present invention is not limited to the scope of the embodiments.
The experimental methods without specifying specific conditions in the following examples were selected according to the conventional methods and conditions, or according to the commercial instructions. The reagents and starting materials used in the present invention are commercially available.
Example 1
1. Characterization and biological function identification of polysaccharide modified bone marrow mesenchymal stem cells
1.1.Ac4ManNAz polysaccharide Effect on expression of sternness markers in rat BMSCs
To assess whether the stem cell characteristics of BMSCs changed after Ac4ManNAz polysaccharide treatment, we examined changes in mesenchymal stem cell markers on the cell surface of BMSCs by flow cytometry. After BMSCs were treated with different concentrations of polysaccharides (0. Mu. Mol/L, 5. Mu. Mol/L, 25. Mu. Mol/L, 50. Mu. Mol/L, 100. Mu. Mol/L), the expression of CD29, CD90, CD34 and CD45 on the surface of stem cells was not significantly different from that of stem cells not treated with polysaccharides (FIG. 1A), both CD29 and CD90 were highly expressed (more than 99%), while CD34 and CD45 were significantly less expressed (less than 5%). Further quantitative analysis also showed that the expression of CD29, CD90, CD34 and CD45 on the surface of BMSCs was not affected after polysaccharide treatment (fig. 1B).
Effect of Ac4ManNAz polysaccharide on rat BMSCs trilineage differentiation
We assessed the effect of polysaccharides on the differentiation capacity of BMSCs by analyzing whether cells have altered osteogenic, chondrogenic, adipogenic differentiation capacity.
1.2.1 osteogenic differentiation:
(1) alkaline phosphatase (ALP) staining: after 7 days of induction in osteogenic induction medium, alkaline phosphatase staining was performed, and the results showed: the BMSCs treated by 25 mu mol/L polysaccharide have no obvious difference in the amount of cells which are positively stained between the BMSCs not treated by polysaccharide, while the blank control group has no obvious positive staining due to the absence of osteogenesis induction medium (FIG. 2A). The results of the quantitative analysis of ALP activity also showed no significant difference between the control group and the experimental group (fig. 2B).
(2) Alizarin red staining: alizarin red staining is carried out after 14 days of osteogenic induction, and results show that the staining intensity between the control group and the experimental group has no significant difference, which indicates that the calcium nodule deposition level between the BMSCs treated by 25 mu mol/L polysaccharide and the two groups of untreated BMSCs after osteogenic induction has no difference, and no obvious calcium deposition is seen in the blank control group (fig. 3A). Alizarin red quantitative staining results also showed no significant difference between the control and experimental groups (fig. 3B).
(3) Real-time quantitative reverse transcription polymerase chain reaction (qRT-PCR): 7 days after osteogenesis induction, transcription levels of osteogenic genes were determined using qRT-PCR, and the assay indexes included alkaline phosphatase (Alp) and osteopontin (Opn). As shown in FIG. 4, the expression of the polysaccharide-treated osteogenic genes was not significantly different from that of the untreated BMSCs after osteogenic induction. The results show that the osteogenic differentiation capacity of BMSCs is not affected after the BMSCs are treated by the polysaccharide.
1.2.2 chondrogenic differentiation:
(1) toluidine blue staining: after 14-16 days of chondrogenic induction medium induction, toluidine blue staining was performed, and the results indicated that there was no significant difference in the amount of positive staining between the control group and the experimental group, while the blank control group did not use chondrogenic induction medium for induction culture and did not see positive staining areas (fig. 5).
1.2.3 adipogenic differentiation
(1) Dyeing with oil red O: oil red O staining is carried out 14-16 days after the adipogenic induction, and the formation of lipid droplets is observed. The oil O staining results indicated that there were a large number of lipid droplets formed between the control and experimental BMSCs after 14 days induction in the lipid induction medium, with no significant difference between the two groups (fig. 6A).
(2) Reverse transcription-polymerase chain reaction (RT-PCR): 7 days after the induction of BMSCs adipogenic differentiation by the adipogenic induction medium, the transcription level of adipogenic related genes Adiponectin and PPar-gamma is determined by PCR. The PCR results show that: after induction in the adipogenic induction medium, the expression of Adiponectin and Ppar-gamma was not significantly different between the control group and the experimental group (fig. 6B and C). The dyeing and PCR results show that the polysaccharide treatment has no obvious influence on the fat-forming differentiation capability of the BMSCs.
1.3 rat BMSCs surface azido groups (N) 3 ) Detection of (2)
To test whether the surface of rat BMSCs is grafted with N azide group 3 Detecting cell surface N by using fluorescent probe with DBCO 3 Expression of (2). The result of fluorescent staining shows that the azide group N is formed after rat BMSCs are cultured in a culture medium containing polysaccharide 3 The BMSCs are successfully grafted on the cell surface, and compared with the common culture medium, the BMSCs cultured by the culture medium containing the polysaccharide show obvious fluorescent signals, but the BMSCs cultured by the common culture medium do not see the fluorescent signals (figure 7). We further determined whether different concentrations of polysaccharide-treated BMSCs exhibited fluorescence of different intensities, and we incubated rat BMSCs with 5. Mu. Mol/L, 25. Mu. Mol/L, 50. Mu. Mol/L and 100. Mu. Mol/L polysaccharide, and the fluorescence staining results showed that the surface fluorescence intensity of BMSCs increased gradually with increasing polysaccharide concentration (FIG. 7).
The fluorescence intensity of the surface of BMSCs after different concentrations of polysaccharide treatment is measured by flow cytometry, the results also show that the fluorescence intensity of the surface of BMSCs is gradually increased along with the increase of the concentration of polysaccharide (0-100 mu mol/L) (FIG. 8A), the fluorescence staining quantitative analysis result shows that the fluorescence intensity of the surface of BMSCs is increased along with the increase of the concentration of polysaccharide, the statistical difference exists among groups, and the fluorescence intensity of the surface is obviously higher than 25 mu mol/L after 50 mu mol/L and 100 mu mol/L polysaccharide treatment (FIG. 8B). However, in combination with the results of CCK-8, the proliferation activity of BMSCs is significantly affected at these two concentrations, so that the cell viability is not affected and a significant fluorescence signal can be detected by using 25. Mu. Mol/L polysaccharide to intervene in the cells in the present study.
2. Characterization and identification of surface of mussel bionic polypeptide-modified titanium implant
2.1XPS and EDS determination of chemical composition of the surface of titanium materials
Synthesizing DBCO modified titanium affinity mussel bionic polypeptide by utilizing an Fmoc solid phase synthesis method, wherein the polypeptide sequence is as follows: ac- (DOPA) -Gly- (DOPA) -Lys [ (PEG 5) - (Mpa) - (Mal-DBCO)]- (DOPA) -Gly- (DOPA). The polypeptide can be reacted with TiO on the surface of a titanium material through DOPA groups 2 And the coordination is carried out, and the titanium material is grafted to the surface of the titanium material. The purity and molecular weight of the polypeptide were determined by HPLC and EI-MS mass spectrometry, and the result showed that the purity of the polypeptide was 97.35%, and that the polypeptide had [ M-H ]]1825.0, consistent with a calculated polypeptide molecular weight of 1825.94 (fig. 9).
In order to clarify the change of the chemical element composition after the polypeptide is grafted on the surface of the titanium material, the XPS spectrum characterizes the surface of the titanium sheet, and the results show that the surface element composition shows an obvious N1s signal enhancement performance after the polypeptide is grafted on the surface of the titanium material, which indicates that the polypeptide is successfully grafted on the surface of the titanium material through coordination bonds after the polypeptide is soaked in the titanium material, and the enhancement of the N element composition is shown (fig. 10A). Furthermore, the quantitative analysis of elements by EDS energy dispersive X-ray spectrometer (FIG. 10B) revealed that the N content was significantly higher after soaking the polypeptide in the titanium plate than before soaking (4.95. + -. 1.00% vs. 8.49. + -. 1.96% in the control group), and the N/Ti atomic ratio was increased from 0.75 to 2.46. The polypeptide is grafted on the surface of the titanium material and then the N element is introduced.
2.2 atomic force microscope detection of titanium sheet surface morphology
In order to evaluate the surface morphology and roughness of the titanium material after the polypeptide is grafted with the titanium material, the surface of the modified titanium material and the surface of the unmodified titanium material are observed by an AFM microscope. As seen in FIG. 11, the surface morphology of the titanium material is regular, and the roughness of the surface of the titanium material after the polypeptide grafting is obviously increased, which indirectly proves that the polypeptide is successfully grafted.
2.3 measurement of hydrophilic/hydrophobic Properties of titanium surface grafted with Polypeptides by Water contact Angle
To evaluate the hydrophilicity and hydrophobicity of the titanium material surface after polypeptide grafting, a water contact angle measurement was performed. As shown in fig. 12, the titanium sheet without grafted polypeptide shows hydrophobicity, the average value of the water contact angle is 45.80 ± 4.67 °, the water contact angle after grafted titanium sheet is significantly reduced, and the average value is 22.60 ± 2.96 °, which shows that the hydrophobicity of the titanium material is changed after grafted polypeptide, the water contact angle is significantly reduced, the hydrophilicity is significantly increased, and the titanium material is beneficial to playing a role.
Example 2
1. Promotion of titanium implant osseointegration based on bio-orthogonal click chemistry
1.BMSC-N 3 Osteogenic differentiation capacity after conjugation to DOPA-DBCO polypeptide modified titanium materials
In order to detect whether the osteogenic differentiation capacity of BMSCs subjected to chemical reaction between cells and the surface of a material is influenced after the BMSCs modified with azide groups are combined with a titanium material modified with DOPA-DBCO polypeptide. BMSCs modified or unmodified by polysaccharide are inoculated on the surfaces of different titanium materials and subjected to osteogenic induced differentiation.
The results of alizarin red staining at day 14 and 21 also showed no significant difference between the calcium nodule deposition positive staining regions of the other groups except the blank control group (fig. 13A), and no significant difference between the alizarin red quantitation results of the groups (fig. 13B). Early bone formation related detection (ALP staining) and late bone matrix mineralization (alizarin red staining) both indicate that BMSCs are captured and bonded on the surface of a titanium material through the reaction of azide groups and DBCO groups on the surface of the titanium material, and the chemical reaction has no influence on the bone formation differentiation capability of the BMSCs.
2.BMSC-N 3 With TiO 2 In vivo study of osteointegration of titanium implants promoted by DOPA-DBCO
Animal experiments, surgical related instruments or other articles are sterilized in advance according to strict aseptic requirements. Through repeated pre-experiments, the exposure of the lateral condyle of the femur is well mastered, and the titanium screw insertion point is determined. After animal experiments were completed and carefully raised for 4 weeks, femoral specimens were collected and subjected to experiments.
2. Titanium screw-bone interface imaging analysis
And (3) carrying out Micro CT examination on the collected femur specimen, evaluating the new bone formation condition on the surface of the titanium screw, and comparing the difference of osseointegration of each group. Polysaccharide-modified BMSCs and polypeptide-modified titanium screw set (BMSCs-N) in the same region of interest (VOI) 3 +TiO 2 DBCO group) each bone parameter was significantly better than the other groups, which showed higher bone volume fraction (BV/TV) and bone surface area density (BS/TV), and lower bone surface area to bone volume ratio (BS/BV), and was statistically different from the other groups. BMSCs-N 3 +TiO 2 The trabecular bone structure around the titanium screw of the DBCO group exhibits optimal osseointegration conditions: the trabecular bone number (Tb.N) mean value is significantly better than the other groups, and all have statistical differences; average trabecular bone thickness (Tb.Th) is superior to other groups except TiO 2 +BMSCs-N 3 Outside the group, there was statistical significance compared to the other groups; while trabecular bone separation (tb.sp) was lower in average than in the other groups, statistically different from the two groups without BMSCs (fig. 14).
The data analysis result of the Micro CT shows that N is 3 The BMSCs are modified by groups and the DBCO polypeptide is modified for titanium materials, and the double modification strategy can obviously improve the osseointegration on the surface of the titanium implant.
3. Histological analysis of titanium screw-bone interface
Histology is an effective method to directly observe the morphology of bone tissue around titanium screw implants, we used toluidine blue staining of hard tissue sections to study the osseointegration around each group of titanium screws, as shown in fig. 15, when the titanium screws were simply screwed in without injecting BMSCs group, the bone tissue formation around the titanium screws was significantly less than that of other groups, while BMSCs-N3+ TiO2-DBCO group had better bone ingrowth than other groups and it was seen that there was continuous bone matrix covering the screw surface. Bone-implant contact (BIC) is an indicator of the quantitative analysis of the extent of osteointegration, BMSCs-N as shown in FIG. 15 3 +TiO 2 The BIC of the DBCO group was significantly better than the other groups, indicating that this group had good osteointegration. The above results show that the biological base is based onOrthogonal click chemical reaction, after BMSCs and titanium materials are doubly modified, azide groups on the surfaces of cells and DBCO groups on the surfaces of the materials are subjected to click chemical reaction, so that more BMSCs can be combined on the surfaces of the materials, the osteogenic differentiation effect of the BMSCs is exerted, and the titanium screw has better osseointegration.
4. Biomechanical testing of titanium screw-bone interface
The anti-extraction force of the titanium screw is detected through a biomechanical test, and the osseointegration condition of the screw and the surrounding bone tissue is evaluated. The more stable the screw is in engagement with the surrounding bone tissue, the stronger the pullout resistance. As shown in FIG. 16, the anti-pullout force of each group containing BMSCs was significantly improved as compared with the titanium screw group alone and the titanium screw-modified group, and BMSCs-N was observed 3 +TiO 2 The DBCO group had the best resistance to pullout, indicating that the addition of BMSCs, which promoted osteodifferentiation around the screw, increased osteointegration strength and resistance to pullout, increased the number of local seed cells of the screw. However, with the simple addition of BMSCs, cells can be difficult to be combined on the surface of the material in a large amount, most of the cells can be lost and can not play a role, and by modifying azide groups on the surface of the BMSCs, the BMSCs-N can be enabled to be subjected to click chemistry reaction 3 Is made of TiO 2 The DBCO is captured and combined on the surface of the material in a large amount, so that the number of seed cells on the surface of the titanium material is obviously increased, and the osseointegration efficiency can be improved. The result of the biomechanical test is consistent with that of a hard tissue slice, and the result shows that the strategy of bioorthogonal click chemical modification can well improve the peripheral osseointegration of the titanium implant, make up for the defect of biological inertia of the titanium material, and prevent the titanium material from loosening and the like to cause the failure of the implantation operation.
The above embodiments are preferred embodiments of the present invention, but the present invention is not limited to the above embodiments, and any other changes, modifications, substitutions, combinations, and simplifications which do not depart from the spirit and principle of the present invention should be construed as equivalents thereof, and all such modifications are intended to be included in the scope of the present invention.
Claims (10)
1.A titanium implant, which is characterized in that azide group N is grafted 3 The bone marrow mesenchymal stem cells and the mussel bionic polypeptide repairCombining the decorated titanium materials to obtain the titanium implant.
2. The titanium implant of claim 1, wherein bone marrow mesenchymal stem cells are captured and bound to the surface of the titanium material by bio-orthogonal click chemistry reaction of azide groups with DBCO groups on the surface of the titanium material.
3. The titanium implant of claim 1, wherein said titanium implant is a titanium screw.
4. A method for promoting osseointegration and/or bone regeneration repair based on bio-orthogonal click chemistry reaction, characterized in that a titanium implant obtained by bio-orthogonal click chemistry reaction is implanted into an animal body to promote osseointegration and/or bone regeneration repair.
5. The process according to claim 4, characterized in that the azide group N is grafted 3 The bone marrow mesenchymal stem cells are combined with the mussel bionic polypeptide modified titanium material to obtain the titanium implant.
6. The method of claim 5, wherein the step of treating the mesenchymal stem cells with the polysaccharide grafts the azide group N on the surface of the mesenchymal stem cells 3 。
7. The method of claim 6, wherein the concentration of the polysaccharide is 0-100 μmol/L.
8. The method of claim 7, wherein the polysaccharide is present at a concentration of 20-30 μmol/L.
9. The method of claim 5, wherein the DBCO-modified mussel biomimetic polypeptide is synthesized using Fmoc solid phase synthesis.
10. The method of claim 9, wherein the mussel biomimetic polypeptide is grafted onto the surface of the titanium material by coordinating with TiO2 on the surface of the titanium through DOPA groups.
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| CN114869911A (en) * | 2022-04-27 | 2022-08-09 | 中山大学·深圳 | Application of PD-1 cell membrane nano vesicle combined stem cell membrane in postoperative treatment of malignant melanoma |
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| US20210145889A1 (en) * | 2018-04-04 | 2021-05-20 | Sigilon Therapeutics, Inc. | Methods, compositions, and implantable elements comprising stem cells |
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