CN104262651A - Preparation and application of auto-platelet-rich plasma thixotropic separating gel - Google Patents
Preparation and application of auto-platelet-rich plasma thixotropic separating gel Download PDFInfo
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- CN104262651A CN104262651A CN201410487483.2A CN201410487483A CN104262651A CN 104262651 A CN104262651 A CN 104262651A CN 201410487483 A CN201410487483 A CN 201410487483A CN 104262651 A CN104262651 A CN 104262651A
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Abstract
The invention relates to preparation and application of auto-platelet-rich plasma thixotropic separating gel, belonging to the fields of high polymer chemistry and biomedicine. The preparation method comprises the following steps: by using acrylate monomer, nano silicon dioxide and silanization activator as raw materials, stirring and heating to react at 60-140 DEG C for 1-4 hours in a nitrogen protective atmosphere; and adding a photoinitiator, irradiating with ultraviolet light to obtain a thixotropic composite high-polymer-base adhesive, blending the thixotropic composite high-polymer-base adhesive with dioctyl phthalate and Span 80, carrying out vacuum degassing and irradiating with cobalt-60 gamma rays. The thixotropic separating gel has excellent thixotropy, biocompatibility and stability, and is suitable for safely, simply and completely extracting auto-platelet-rich plasma and other bioactive substances in biomedicine.
Description
Technical field
The present invention relates to the preparations and applicatio of autologous platelet rich plasma thixotroping separating gel, belong to polymer chemistry and biomedical sector.
Background technology
Autologous platelet rich plasma (Platelet-rich plasma, PRP) is the platelet concentrate that autologous blood produces through separation, and autologous PRP derives from autologous, and safety performance is good, can fundamentally eliminate the problem such as pathophoresis and immunological rejection.A large amount of clinical study report, PRP contains a large amount of various platelet derived cell growth factors, has the effect promoting that Various Tissues is repaired, can promote the reparation of bone and soft tissue.PRP promotes that bone defect healing may mainly based on following two kinds of mechanism: one is that PRP activates under activator effect is rich platelet gel (Platelet-rich gel, PRG), wherein containing a large amount of Fibrinogen, energy wound closure, promotes tissue contracts and healing, another kind is that in polymerization process, thrombocyte shrinks, retting conditions discharges multiple high density somatomedin, comprise Thr6 PDGF BB (PDGF), transforming growth factor (TGF-β), vascular endothelial growth factor (VEGF) and Urogastron (EGF) etc., PDGF and TGF-β is the most important thing is in these cell growth factors, the effect of PRP is that multiple somatomedin forms a series of signal transduction by different approaches, the expression of activation corresponding gene has come, play an important role in tissue repair with reconstruction: PDGF can stimulate the mitotic division of bone marrow stroma stem cell, increase osteoclast number, also can the mitotic division of stimulating endothelial cell, promote the angiogenesis of graft area, and increase the ability of collagen protein synthesis, TGF-β then can promote that preosteoblast is bred and suppresses bone resorption.In PRG, other somatomedins also have good promotion bone and the effect of repair of cartilage and regeneration simultaneously.Thus PRP is used for clinical treatment osteoarthritis by external many developed countries, and by the autologous PRP of joint cavity injection, treated effect reaches more than 80%.So autologous PRP has important using value and wide DEVELOPMENT PROSPECT in biomedicine.
But conventional P RP preparation method is separated by manual absorption mode to obtain PRP.The method is centrifugal preparation PRP in open system, and shifts through multiple container, and process is more loaded down with trivial details, easily by outside contamination; The middle thrombocyte that different centrifugal number of times, centrifugal force and centrifugation time make and somatomedin concentration and activity also different; And manual absorption mode easily makes platelet activation, affect effective acquisition of somatomedin.So conventional P RP preparation method can not guarantee PRP safety, effectively extract, thus have impact on the Application and Development of autologous PRP at biomedical sector.
Summary of the invention
The present invention is directed to the preparations and applicatio that the shortcoming and defect existed in prior art provides a kind of autologous platelet rich plasma thixotroping separating gel.It has good stability, is applicable to prepare the extraction of the high autologous platelet rich plasma thixotroping separating gel of security requirement.
For solving the problems of the technologies described above, the technical solution used in the present invention is:
Autologous platelet rich plasma thixotroping separating gel, is characterized in that: it is with acrylate monomer, nano silicon and silanization promoting agent for raw material, stirring heating reaction under nitrogen protection, 1 ~ 4 hour reaction times, temperature of reaction 60 ~ 140 DEG C; Then light trigger is added, ultraviolet light irradiation, obtained thixotropy composite high-molecular base glue, then thixotropy composite high-molecular base glue and dioctyl phthalate (DOP) and span80 are carried out blended, vacuum defoamation, then obtains autologous platelet rich plasma thixotroping separating gel through cobalt-60γray irradiation.
By such scheme, be filled in after bottom traditional vacuum separator tube after described thixotroping separating gel vacuum defoamation, then prepare autologous platelet rich plasma thixotroping separating gel through cobalt-60γray irradiation.
By such scheme, described acrylate monomer is the mixing of propyl acrylate and Hydroxyethyl acrylate, or be the mixing of propyl acrylate, Hydroxyethyl acrylate and isoamyl acrylate, described acrylate monomer be propyl acrylate and Hydroxyethyl acrylate time, the mass ratio of two kinds is 1:0.02-0.04; During for propyl acrylate, Hydroxyethyl acrylate and isoamyl acrylate, the mass ratio of three is 1:0.03-0.05:0.1-0.5.
By such scheme, the particle diameter of described nano silicon is 30-50nm, and its consumption is the 1-15wt% of acrylate monomer total mass.
By such scheme, described silanization promoting agent is dodecyl trimethoxy siloxane, and its consumption is the 5-20wt% of nano silicon quality.
By such scheme, the light trigger of described photopolymerization reaction is gorgeous solid good (Irgacure) 2595, and its consumption is 0.10 ~ 0.30wt% of acrylate monomer total mass.
By such scheme, described ultraviolet light irradiation wavelength is 365nm, and irradiation power is 30mWcm
-2, irradiation time is 3-20min.
By such scheme, described dioctyl phthalate (DOP) and span80 consumption are the 0.5-5.0wt% of thixotropy composite high-molecular base colloid amount.
By such scheme, described cobalt-60γray irradiation dose is 20 ~ 30kGy.
By such scheme, the apparent viscosity of described autologous platelet rich plasma thixotroping separating gel is 1 ~ 20 × 10
4mPas(rotational viscosimeter test gained, probe temperature 25 DEG C, test rotating speed 0.3rpm), proportion is 1.09 ~ 1.15.
The application of autologous platelet rich plasma thixotroping separating gel in preparing for autologous platelet rich plasma, its application method is: have in the traditional vacuum separator tube of autologous platelet rich plasma thixotroping separating gel in underfill, add after anticoagulant solution vacuumizes, introduce the autologous whole blood preparing autologous platelet rich plasma to be extracted, then autologous platelet rich plasma is prepared in centrifugation.
Beneficial effect of the present invention:
1) the autologous platelet rich plasma thixotroping separating gel provided by the invention polyacrylate composite nano gel that is the physical crosslinkings such as a kind of existing hydrogen bond, has chemical bond crosslinked again, it has good stability, contact with anticoagulant solution, the stripping of gel-free composition.There is excellent thixotropy and biocompatibility simultaneously.Be applicable to biomedical safe, easy, fully extract the autologous platelet rich plasma high to security requirement.Although there is the report of serum separation gel at present, existing serum separation gel is all with the plural gel of the physical crosslinkings such as hydrogen bond, less stable, can not be applicable to the separation preparation of PRP.
2) the present invention is by utilizing the thixotropy of autologous platelet rich plasma thixotroping separating gel and PRP and blood cell specific gravity difference, can realize effective separation of PRP, isolation and extraction under the influence of centrifugal force.Detect through platelet count, compared with whole blood, in this PRP, PC increases by more than 3 times.
3) the present invention can make whole PRP extract operation all to complete in an airtight sterile test tube is as traditional vacuum separator tube, and operation efficiency improves greatly, for biomedicine extract autologous PRP provide one very safe, stablize and method easily.
Embodiment
By the following examples summary of the invention of the present invention is elaborated.
Embodiment 1
100g propyl acrylate, 2.0g Hydroxyethyl acrylate, 6.0g nano silicon and 1.0g silanization promoting agent are put in 250ml four-hole bottle, load onto thermometer, mechanical stirrer, still head and condensate collection device, import high pure nitrogen, open water of condensation, and start slowly to be heated to 60 ~ 80 DEG C.Stirring reaction 1 hour under nitrogen protection.Add gorgeous solid good (Irgacure) 2595 of light trigger 0.2g again, with UV-light (365nm, 30mWcm
-2) irradiation 3 minutes, the obtained thixotropy composite high-molecular base glue of reaction.Then carry out blended with 2.0g dioctyl phthalate (DOP) and 0.6g span80, vacuum defoamation, be filled in bottom traditional vacuum separator tube, through cobalt-60γray irradiation (30kGy), prepare thixotropy composite high-molecular gel.
The apparent viscosity of this gel is 1.885 × 10
5mPa ﹒ s(rotational viscosimeter test gained, probe temperature 25 DEG C, rotating speed 0.3rpm), thixotropy index 3.61, proportion 1.13.Get it to be about 2g and to be filled in bottom traditional vacuum separator tube, add after anticoagulant solution vacuumizes, introduce the autologous whole blood 10 milliliters preparing autologous platelet rich plasma to be extracted, then autologous platelet rich plasma is prepared in centrifugation.Detect through platelet count, compared with whole blood, in this PRP, PC increases by 3.12 times.
Embodiment 2
100g propyl acrylate, 4.0g Hydroxyethyl acrylate, 4.0g nano silicon and 0.6g silanization promoting agent are put in 250ml four-hole bottle, load onto thermometer, mechanical stirrer, still head and condensate collection device, import high pure nitrogen, open water of condensation, and start slowly to be heated to 70 ~ 90 DEG C.Stirring reaction 1.5 hours under nitrogen protection.Add gorgeous solid good (Irgacure) 2595 of light trigger 0.3g again, with UV-light (365nm, 30mWcm
-2) irradiation 5 minutes, the obtained thixotropy composite high-molecular base glue of reaction.Then carry out blended with 3.5g dioctyl phthalate (DOP) and 0.8g span80.Be filled in bottom traditional vacuum separator tube, vacuum defoamation, cobalt-60γray irradiation (20kGy), prepare thixotropy composite high-molecular gel.
The apparent viscosity of this gel is 1.045 × 10
5mPa ﹒ s(rotational viscosimeter test gained, temperature 25 DEG C, rotating speed 0.3rpm), thixotropy index 3.45, proportion 1.09.Get the traditional vacuum separator tube that above-mentioned underfill has thixotropy composite high-molecular gel, add after anticoagulant solution vacuumizes wherein, introduce the autologous whole blood preparing autologous platelet rich plasma to be extracted, then autologous platelet rich plasma is prepared in centrifugation.Detect through platelet count, compared with whole blood, in this PRP, PC increases by 3.54 times.
Embodiment 3
100g propyl acrylate, 3.0g Hydroxyethyl acrylate, 20g isoamyl acrylate, 7.5g nano silicon and 1.1g silanization promoting agent are put in 250ml four-hole bottle, load onto thermometer, mechanical stirrer, still head and condensate collection device, import high pure nitrogen, open water of condensation, and start slowly to be heated to 80 ~ 90 DEG C.Stirring reaction 2 hours under nitrogen protection.Add gorgeous solid good (Irgacure) 2595 of light trigger 0.3g again, with UV-light (365nm, 30mWcm
-2) irradiation 5 minutes, the obtained thixotropy composite high-molecular base glue of reaction.Then carry out blended with 5.0g dioctyl phthalate (DOP) and 0.6g span80.Vacuum defoamation, prepares thixotropy composite high-molecular gel through cobalt-60γray irradiation (25kGy).
The apparent viscosity of gained gel is 1.132 × 10
5mPa ﹒ s(temperature 25 DEG C, rotating speed 0.3rpm), thixotropy index 3.70, proportion 1.10.Get it to be about 2g and to be filled in bottom traditional vacuum separator tube, add after anticoagulant solution vacuumizes, introduce the autologous whole blood 10 milliliters preparing autologous platelet rich plasma to be extracted, then autologous platelet rich plasma is prepared in centrifugation.Detect through platelet count, compared with whole blood, in this PRP, PC increases by 3.68 times.
Embodiment 4
100g propyl acrylate, 4.0g Hydroxyethyl acrylate, 6.0g nano silicon and 1.0g silanization promoting agent are put in 250ml four-hole bottle, load onto thermometer, mechanical stirrer, still head and condensate collection device, import high pure nitrogen, open water of condensation, and start slowly to be heated to 60 ~ 80 DEG C.Stirring reaction 1 hour under nitrogen protection.Add gorgeous solid good (Irgacure) 2595 of light trigger 0.2g again, with ultraviolet light irradiation 3 minutes, the obtained thixotropy composite high-molecular base glue of reaction.Then carry out blended with 4.0g dioctyl phthalate (DOP) and 0.5g span80, vacuum defoamation, prepare thixotropy composite high-molecular gel through cobalt-60γray irradiation (30kGy).
The apparent viscosity of gained gel is 1.625 × 10
5mPa ﹒ s(temperature 25 DEG C, rotating speed 0.3rpm), thixotropy index 3.35, proportion 1.10.Get it to be about 2g and to be filled in bottom traditional vacuum separator tube, add after anticoagulant solution vacuumizes, introduce the autologous whole blood 10 milliliters preparing autologous platelet rich plasma to be extracted, then autologous platelet rich plasma is prepared in centrifugation.Detect through platelet count, compared with whole blood, in this PRP, PC increases by 3.18 times.
Embodiment 5
100g propyl acrylate, 3.0g Hydroxyethyl acrylate, 20g isoamyl acrylate, 8.5g nano silicon and 1.4g silanization promoting agent are put in 250ml four-hole bottle, load onto thermometer, mechanical stirrer, still head and condensate collection device, import high pure nitrogen, open water of condensation, and start slowly to be heated to 80 ~ 90 DEG C.Stirring reaction 2 hours under nitrogen protection.Add gorgeous solid good (Irgacure) 2595 of light trigger 0.3g again, with ultraviolet lighting 5 minutes, the obtained thixotropy composite high-molecular base glue of reaction.Then carry out blended with 3.0g dioctyl phthalate (DOP) and 1.0g span80.Vacuum defoamation, prepares thixotropy composite high-molecular gel through cobalt-60γray irradiation (25kGy).
The apparent viscosity of gained gel is 1.832 × 10
5mPa ﹒ s(temperature 25 DEG C, rotating speed 0.3rpm), thixotropy index 3.75, proportion 1.14.Get it to be about 2g and to be filled in bottom traditional vacuum separator tube, add after anticoagulant solution vacuumizes, introduce the autologous whole blood 10 milliliters preparing autologous platelet rich plasma to be extracted, then autologous platelet rich plasma is prepared in centrifugation.Detect through platelet count, compared with whole blood, in this PRP, PC increases by 3.78 times.
Claims (10)
1. autologous platelet rich plasma thixotroping separating gel, is characterized in that: it is with acrylate monomer, nano silicon and silanization promoting agent for raw material, stirring heating reaction under nitrogen protection, 1 ~ 4 hour reaction times, temperature of reaction 60 ~ 140 DEG C; Then light trigger is added, ultraviolet light irradiation, obtained thixotropy composite high-molecular base glue, then thixotropy composite high-molecular base glue and dioctyl phthalate (DOP) and span80 are carried out blended, vacuum defoamation, then obtains autologous platelet rich plasma thixotroping separating gel through cobalt-60γray irradiation.
2. autologous platelet rich plasma thixotroping separating gel according to claim 1, it is characterized in that: be filled in after bottom traditional vacuum separator tube after described thixotroping separating gel vacuum defoamation, then prepare autologous platelet rich plasma thixotroping separating gel through cobalt-60γray irradiation; Described cobalt-60γray irradiation dose is 20 ~ 30kGy.
3. autologous platelet rich plasma thixotroping separating gel according to claim 1, it is characterized in that: described acrylate monomer is the mixing of propyl acrylate and Hydroxyethyl acrylate, or be the mixing of propyl acrylate, Hydroxyethyl acrylate and isoamyl acrylate, described acrylate monomer be propyl acrylate and Hydroxyethyl acrylate time, the mass ratio of two kinds is 1:0.02-0.04; During for propyl acrylate, Hydroxyethyl acrylate and isoamyl acrylate, the mass ratio of three is 1:0.03-0.05:0.1-0.5.
4. autologous platelet rich plasma thixotroping separating gel according to claim 1, is characterized in that: the particle diameter of described nano silicon is 30-50nm, and its consumption is the 1-15wt% of acrylate monomer total mass.
5. autologous platelet rich plasma thixotroping separating gel according to claim 1, it is characterized in that: described silanization promoting agent is dodecyl trimethoxy siloxane, its consumption is the 5-20wt% of nano silicon quality.
6. autologous platelet rich plasma thixotroping separating gel according to claim 1, is characterized in that: the light trigger of described photopolymerization reaction is Irgacure2595, and its consumption is 0.10 ~ 0.30wt% of acrylate monomer total mass.
7. autologous platelet rich plasma thixotroping separating gel according to claim 1, it is characterized in that: described ultraviolet light irradiation wavelength is 365nm, irradiation power is 30mWcm
-2, irradiation time is 3-20min.
8. autologous platelet rich plasma thixotroping separating gel according to claim 1, is characterized in that: described dioctyl phthalate (DOP) and span80 consumption are the 0.5-5.0wt% of thixotropy composite high-molecular base colloid amount.
9. autologous platelet rich plasma thixotroping separating gel according to claim 1, is characterized in that: the apparent viscosity of described autologous platelet rich plasma thixotroping separating gel is 1 ~ 20 × 10
4mPas, proportion is 1.09 ~ 1.15.
10. the application of the autologous platelet rich plasma thixotroping separating gel according to any one of claim 1-9 in preparing for autologous platelet rich plasma, its application method is: have in the traditional vacuum separator tube of autologous platelet rich plasma thixotroping separating gel in underfill, add after anticoagulant solution vacuumizes, introduce the autologous whole blood preparing autologous platelet rich plasma to be extracted, then autologous platelet rich plasma is prepared in centrifugation.
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| CN106366426A (en) * | 2016-08-30 | 2017-02-01 | 成都瑞琦科技实业股份有限公司 | Separation gel system for platelet-rich plasma (PRP) extraction and purification and preparation method thereof |
| CN107353364A (en) * | 2017-06-27 | 2017-11-17 | 湖北博成生物科技有限公司 | A kind of hydrolysis blood separating colloid and preparation method thereof |
| CN111468204A (en) * | 2020-06-08 | 2020-07-31 | 珠海朗泰生物科技有限公司 | Platelet-rich plasma preparation tube with controllable components and preparation method thereof |
| CN113717320A (en) * | 2020-05-25 | 2021-11-30 | 华熙生物科技股份有限公司 | Preparation method of platelet-rich plasma separation gel, obtained product and application |
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