TWI448312B - An implant of freeze - dried platelets and its coating method - Google Patents

Description

Implanted freeze-dried platelet implant and coating method thereof

The present invention relates to an implant coated with protected freeze-dried platelets and a method of coating the same.

A platelet-rich plasma (hereinafter referred to as PRP) or a related art thereof is known in the art, and is disclosed in, for example, U.S. Patent No. 7,769, 922, 654, 554, pp. PRP is coated with implant technology, but the PRP or its related components involved in these technologies are not protected by protective steps. Therefore, the problem of rapid decay of effective price must be used immediately after blood collection, or cryopreservation. Shortcomings of short shelf life. In the lyophilization method, the protected platelets are coated on the implant, and the titer of the platelet cells can be stored at room temperature for one year, and there is still no significant change, and the coated implants are protected in the coating. After freeze-dried platelets, even if stored at room temperature, the effective period can be as high as 3 years or more.

The present invention provides an implant coated with freeze-dried platelets, wherein the freeze-dried platelets are protected freeze-dried platelets.

The present invention provides a method of coating freeze-dried platelets on an implant, wherein the freeze-dried platelets are protected freeze-dried platelets.

The invention discloses a method for coating freeze-dried platelets on an implant, Protecting the freeze-dried platelets on the implant, the coating method comprising: a coating step of coating the protected platelet solution on the surface of the implant; and a lyophilization step of coating the platelet solution The implant is lyophilized to freeze the platelet solution coated on the surface of the implant into lyophilized platelets.

The implant may be any conventional orthopedic implant (for example, a nail, a bone filler, a cage, etc.) or a dental implant (such as a root).

The above-mentioned protected platelet solution refers to a protected platelet thick solution or a diluted solution thereof, or a platelet solution prepared by reconstituting the protected platelet dry powder, and the platelet made by using the protected platelet dry powder reconstituted water. A solution, or a protected platelet dilution solution is preferred, and a protected platelet dilution solution is more preferred.

The above-mentioned so-called protected platelet thick liquid refers to the addition of a specific reagent in a blood or blood preparation (preferably using PRP), so that the platelet cells can not substantially rupture or substantially reduce the platelet cells in the process of separating the platelets. The rupture ratio, and then the plasma layer is separated to obtain a platelet thick solution. The protection method may be a technique known to the manufacturer (for example, see Taiwan No. I300806, I270375 Announced Patent, Taiwan No. 201004659, 200526680, Patent No. 7,659,052, hereinafter referred to as reference technology), or use adenosine diphosphate ( Adenosine diphosphate (hereinafter referred to as ADP) protection technology (one of the important techniques of the present invention, see the following preliminary examples), a platelet thick solution prepared by the ADP protection program, or a dilution obtained by diluting the platelet thick solution a solution, or a reconstituted platelet dry powder prepared from the platelet thick solution or a diluted solution thereof A platelet solution is preferred because ADP is easy to obtain, the quality is stable, and the method of control is relatively easy.

Here, the process of preparing a platelet thick liquid by the ADP protection program is: adding 0.1 to 10 mM ADP solution to the thick plasma of mammalian platelets, and stirring uniformly to protect the platelets, wherein the ADP dosage is platelet-rich plasma. For every 1 million platelet cells, 0.04 to 4 Nemo ADP was used; the protected platelet thick plasma was removed from the plasma layer to obtain a protected platelet thick solution.

The concentration of the above ADP solution is preferably 0.2 to 5 mM, more preferably 0.5 to 2 mM.

The above ADP solution is preferably used in an amount of 0.1 to 2 nanomolar ADP per plate of platelet-rich plasma per 1 million platelet cells, and more preferably 0.2 to 1 nemoex ADP.

The above-mentioned protected platelet diluted solution refers to a solution obtained by diluting the above-mentioned protected platelet thick solution with water, an aqueous solution (for example, physiological saline).

The above-mentioned protected dry powder of platelets refers to a dry powder of platelets obtained by freeze-drying the above-mentioned protected platelet thick solution or the protected platelet diluted solution. The so-called "platelet solution prepared by reconstituting the protected platelet dry powder" means a solution formed by dissolving the dried dry powder of the protected platelet such as water or an aqueous solution (for example, physiological saline). The method for protecting the dry powder of the platelet is prepared by drying and lyophilizing the platelet thick liquid prepared by the manufacturer's conventional protection technology; or the platelet thick liquid prepared by using the ADP protection program is dried, Made by lyophilization; good.

The above protected platelet solution has a concentration of 50K~20M/μL (containing 50,000 to 20 million platelet cells per ml), preferably 100K~10M/mL, and 200K~5000K/μL. good. Those skilled in the art know that a concentration of less than 50 K/μL is also feasible, but will make the freeze-drying efficiency worse (it must be coated multiple times, lyophilized, and each freeze-drying time is long), so it is feasible but not suitable; Concentrations above 20M/μL are also possible, but are more suitable for special conditions, such as for soaking coating or coating procedures, while coating procedures are generally not suitable for mass production; if used for more suitable mass production , will cause a considerable number of platelets to escape the air.

The protected platelet solution may be added with any active ingredient having a positive effect (for example, an anti-inflammatory component, an analgesic component, a nutrient component, and/or an ingredient which enhances absorption), or a component having substantially no negative effect or side effect (for example, color) In order to make the implant have a specific color, as a label for different products).

The step of coating the protected platelet solution on the surface of the implant may be carried out by a conventional method, for example, spraying the protected platelet solution on the surface of the implant, applying the protected platelet solution to the surface of the implant, Or immerse the implant in the protected platelet solution before taking it out. In general, spraying or coating is preferred, and spraying is preferred; however, under special conditions, such as implants having pores, the protected platelets must be deeply and evenly coated inside the implant. In the case of pores, it is preferred to use a soak coating.

The above platelet sources may be heterologous, homologous or autologous platelets, based on the user's heart. Considering the amount of the platelets, allogeneic platelets or autologous platelets are preferred, and autologous platelets are more preferred. However, based on the source and efficacy considerations, it is preferable to use heterogeneous platelets or allogeneic platelets, and to use heterogeneous (mammalian) platelets as the best, and to use platelets of cattle, pigs or sheep as the best.

In general, platelet-derived growth factor (PDGF) is about 40-200 pg/mL, which has the effect of promoting healing. See Vogt., et al., Determination of endogenous growth factors in human wound healing. Wound Repair Regeneration , 2004, 12 (4): p. 485-492. The effectiveness of the implanted freeze-dried platelet implants of the present invention may be related to the long-term activity of PDGF of platelets, but its exact relevance remains to be confirmed.

The invention also discloses an implant coated with protected freeze-dried platelets, which is based on an implant and coated with protected freeze-dried platelets, wherein the amount of the freeze-dried platelets is: The coated area is coated with 20M to 10000M platelets per square centimeter, preferably 100M to 5000 M. It is not infeasible that the platelet count is higher than the number of platelets per square centimeter of 10000M, but the process will be complicated and complicated. The number of platelets below 20M per square centimeter will cause the release of PDGF to be too low, resulting in curative effect. Not at all.

The above described implants, protected platelet solutions, coatings, lyophilization, and the like are all defined as described above.

To further illustrate the teachings of the present invention, under:

Preliminary Example 1:

The patient was bled with a blood cell separator (MCS System 9000, http://www.haemonetics.com/) to obtain 36.7 mL of autologous PRP.

In a 1.5 mL microcentrifuge tube, add 0.2 mL of physiological saline and 0.2 mL of PRP, and mix well; use a fully automated blood analyzer (Sysmex Corporation KX-21 Automated Hematology Analyzer, see http://www.blockscientific.com) /) measured platelet count cells 476 K / μ L (microliters 476,000 platelets per cell), i.e. cells original PRP platelet number was 952 K / μL, confirmed that meet quality requirements.

With the remaining 36.5 mL of PRP, add 22 mL of a physiological saline solution containing 1 mM ADP (ie 1 mM ADP per liter), ie ADP dosage 0.633 nmole/Mega (0.633 nm per 1 million platelet cells) Moore's ADP), after shaking for 30 minutes, centrifuge at 4000 rpm for 7 minutes, remove the plasma layer, add reverse osmosis water to 35 mL and mix well, and dispense into 35 bottles (1 mL per bottle) Platelet solution. 35 bottles of platelet solution were sent to a -40 ° C freezer (ultra-low temperature horizontal freezer, RS-CF 330LT, Ruey Shing Refrigeration Equipment Co., Ltd.), frozen for 24 hours, and then freeze-dried The machine (FD2-6P-M, Jinming Industrial Co., Ltd. - KINGMECH CO., LTD) was freeze-dried (-40 ° C, 10 Pa) for 36 hours to obtain 35 bottles of platelet dry powder, each containing about one billion platelets.

One of the bottles (Sample 1) was immediately rehydrated according to the following method, and the platelet count and PDGF titer were determined; the remaining 34 bottles were immediately vacuum-packed and externally sterilized. Stored at room temperature for one week (sample 2), two weeks later (sample 3), four weeks later (sample 4), eight weeks later (sample 5), twelve weeks later (sample 6), after sixteen weeks (sample 7) Each bottle of sterilized platelet dry powder was taken, and rehydrated according to the following method, and the platelet count and PDGF titer were measured.

Rehydration procedure: Re-watering samples 1-7 with 4 mL reverse osmosis water.

Platelet count: 0.4 mL reconstituted solution were taken samples 1 to 7, in a fully automatic blood analyzer, respectively platelet number was measured obtaining 216,223,208,219,202,205,201 K / μ L.

Determination of PDGF titer: 1 mL of sample 1~7 of the reconstituted solution was diluted 40 times, and then colorimetrically analyzed by Enzyme-linked immunoassay, using a filter analyzer (Thermo Fisher Scientific Multiskan) FC Filter-based microplate photometer, see http://www.ninolab.se/fileadmin/Ninolab/pdf/thermolabsystems/MultiskanFC_Brochure_0308-01_LR.pdf) Analysis of PDGF titer, the resulting titer multiplied by 40 times, respectively, 5016, 4985 4968, 4902, 4803, 4852, 4785 pg/mL.

The experimental data showed that the platelet count and PDGF titer in the platelet dry powder were quite stable. The platelet count at the 16th week was 93% (201/216) of the original platelet count; the PDGF titer at the 16th week was the original PDGF titer. 95.4% (4785/5016).

Example 1:

The reconstituted aqueous solutions 2, 3, 6, and 7 of Preparation Example 1 were mixed and mixed into a mixed aqueous solution having an average PDGF titer of 4,8 pg/mL.

A porous block-shaped bone filling block (hereinafter referred to as HA block) A, B, and C having a weight of hydroxylapatite (hereinafter referred to as HA) shown in Table 1 was immersed in the above mixed aqueous solution, and evacuated. Continue to soak for 10 minutes, take out the HA block, transfer it to the -40 ° C freezer, freeze for 24 hours, weigh it (see Table 1), and then freeze-dry the HA block A, B, C for 36 hours with a freeze dryer. HA blocks A, B, and C which are coated with freeze-dried platelets.

On the day of preparation, soak the HA block A with 5 mL of reverse osmosis water, soak it for about 30 minutes after vacuuming, and filter it out; repeat the soaking and drying steps for 6 times; concentrate the 7 times of the soaked solution and add water to Dilute to 40 mL. After the dilution (no longer diluted) was developed by enzyme immunoassay, the PDGF titer was analyzed by a filter analyzer as shown in Table 1.

After storage for four weeks at room temperature, HA block B was treated in the same manner, and the PDGF titer was measured as shown in Table 1. After storage for eight weeks at room temperature, HA block C was treated in the same manner, and the PDGF titer was measured as shown in Table 1.

The normalized platelet count and PDGF titer of HA blocks A, B, and C are normalized, and the normalization formula is as follows. The results are shown in Table 1: N=N _j *40/W _j

Wherein N is the normalized PDGF titer; N _j represents the measured value of PDGF titer of HA block A, B, and C, respectively, see Table 1; W _j is the weight of platelet solution covered by HA block A, B, C See Table 1; 40 because the weight of 40 mL is 40 g.

The experimental data showed that the PDGF titers of HA block A (0 weeks), B (4 weeks), and C (8 weeks) were quite consistent with the PDGF titers of the same period (0, 4, and 8 weeks) of the preliminary examples. The ratios were 0.970 (4753/4898), 0.989 (4845/4898), and 0.975 (4776/4898), respectively.

Preliminary Example 2:

PRP (25 mL) was obtained from the blood bank. First, according to the method of the preliminary example 1, 0.2 mL of physiological saline and 0.2 mL of PRP were added to a 1.5 mL small centrifuge tube, and the mixture was thoroughly mixed and then measured by a fully automatic blood analyzer. The number of platelet cells was 712 K/μL (712,000 platelets per microliter), that is, the number of platelets in the original PRP was 1424 K/μL.

14 mL of the above PRP was taken, and the ADP protection procedure of Example 1 was carried out in accordance with the ADP concentration and amount of Table 2. After shaking for 30 minutes, centrifuge and The plasma layer was removed to obtain a platelet layer (hereinafter referred to as PLT). Add reverse osmosis water to 20 mL and mix well to form a platelet solution. Dispense the platelet solution into 10 bottles (2 mL per bottle), then transfer to a -40 ° C freezer, freeze for 24 hours, then freeze. The dryer was freeze-dried (-40 ° C, 10 Pa) for 36 hours to obtain 10 bottles of platelet dry powder, each containing about 2 billion platelets.

Take a bottle of platelet dry powder, immediately rehydrate according to the method described in the preliminary example 1, take 0.5 mL of the reconstituted aqueous solution, dilute 80 times, and then color by enzyme immunoassay, and then analyze the PDGF titer with a filter analyzer. After multiplying the potency by 80 times, the value is 11.42 ng/mL.

Preliminary Examples 3~8:

Platelet dry powder was prepared in the same manner as in Preparation Example 2 except that the ADP concentration and amount were different, see Table 2. The PDGF titer was determined in the same manner as in the preliminary Example 2, and the results are shown in Table 2.

Embodiments 2 to 8:

The dry powders of the preliminary examples 2 to 8 were rehydrated with 4 mL of reverse osmosis water to form a reconstituted aqueous solution 2 to 8 (respectively aqueous solutions of the preliminary examples 2 to 8 respectively).

Seven titanium-based cervical cages (see Table 1 for weight) were immersed in the reconstituted aqueous solution for 2 to 8 hours each. After taking out, it was sent to a -40 ° C freezer, frozen for 24 hours, taken out and weighed, and the weight is shown in Table 3. Then, it was sent to a freeze dryer for freeze-drying for 36 hours to obtain a titanium-based cervical vertebra cage 2~8 coated with freeze-dried platelets.

On the day of preparation, soak the titanium-based cervical vertebra cage with 5 mL of reverse osmosis water for 2~8 for about 30 minutes, and filter it out; repeat the soaking and drying steps for 6 times; concentrate the 7 times of the soaked solution and add water to Dilute to 40 mL. After the dilution (no longer diluted) was developed by enzyme immunoassay, the PDGF titer was analyzed by a filter analyzer as shown in Table 3.

The experimental data showed that the PDGF titer of the titanium-based cervical cage 2~8 was quite consistent with the PDGF titer of the preliminary examples 2-8, and the ratios were 0.928, 0.972, 0.966, 0.982, 0.946, 0.953, 0.943, respectively.

Preparation Example 9:

The platelet dry powder was prepared in the same manner as in the preliminary Example 2, and the ADP protection procedure of Example 1 was carried out in the ADP concentration and amount of Table 4. After shaking for 30 minutes, the plasma layer was centrifuged and removed to obtain PLT. Reverse osmosis Dilute the water to 20 mL and mix well to form a platelet solution. The platelet solution is divided into 5 bottles (4 mL per bottle), then transferred to a -40 ° C freezer, frozen for 24 hours, and then dried in a freeze dryer. Freeze-drying (-40 ° C, 10 Pa), 36 hours, gave 5 bottles of dry powder of platelets, each containing about 4 billion platelets.

Take a bottle of platelet dry powder, immediately rehydrate according to the method described in the preliminary example 1, take 0.5 mL of the reconstituted aqueous solution, dilute 160 times, and then color by enzyme immunoassay, and then analyze the PDGF titer with a filter analyzer. After multiplying the titer by 160 times, the value is 21.05 ng/mL.

Preliminary Examples 10~15:

Platelet dry powder was prepared in the same manner as in Preparation Example 9, except that the ADP concentration and amount were different, see Table 4. The PDGF titer was determined in the same manner as in the preliminary Example 2, and the results are shown in Table 4.

Examples 9-15:

The dry powders of Examples 9 to 15 were reconstituted with 4 mL of reverse osmosis water, respectively, to form a reconstituted aqueous solution of 9 to 15 (reactive aqueous solutions of Preparation Examples 9 to 15, respectively).

The reconstituted aqueous solutions 9-15 were applied to the nails 9-15 (weights are shown in Table 5). It was sent to a -40 ° C freezer, frozen for 24 hours, taken out and weighed, the weight is shown in Table 5. Then, it was sent to a freeze dryer for freeze-drying for 36 hours to obtain bone nails 9 to 15 which were coated with freeze-dried platelets.

On the day of preparation, soak the bone nails in 10 mL of reverse osmosis water for 9 to 15 minutes, filter dry; repeat the soaking and drying steps 6 times; concentrate the 7 times of the soaked solution, add water to dilute to 80 mL. After the dilution of the dilution (no longer diluted) by enzyme immunoassay, the PDGF titer was analyzed by a filter analyzer as shown in Table 5. The normalization formula is as follows: N=N _j *80/W _j

Wherein N is the normalized PDGF titer; N _j represents the measured value of the PDGF titer of the nail, respectively, see Table 5; W _j is the weight of the platelet solution coated by the nail, see Table 5; 80 is because 80 The weight of mL is 80 g.

The experimental data showed that the PDGF titers of bone nails 9-15 were quite consistent with the PDGF titers of the same period of the previous examples 9-15, and the ratios were 0.946, 0.962, 0.957, 0.991, 0.972, 0.928, 0.954, respectively.

Preliminary Example 16:

From a blood cell separator, 620 mL of heterogeneous (porcine) PRP was isolated from approximately 7 liters of pig blood. In a 1.5 mL small centrifuge tube, add 0.8 mL of Physiology The saline solution and 0.2 mL of PRP were mixed well and the number of platelets was 574 K/μL measured by a fully automatic blood analyzer, that is, the number of platelets of the original PRP was 2.87 M/μL (2.77 million per microliter). Platelets).

Take 21 mL of pig PRP and add 30 mL of physiological saline solution containing 1 mM ADP (equivalent to 0.50 nmole ADP/Mega platelets). After shaking for 30 minutes, the plasma layer was centrifuged and removed to obtain PLT. Add reverse osmosis water to 20 ml, mix quickly and form a platelet solution, and dispense into 20 bottles of platelet solution (1 mL per bottle). The platelet solution was sent to a -40 ° C freezer, frozen for 24 hours, and then freeze-dried (-40 ° C, 10 Pa) in a freeze dryer for 36 hours to obtain 20 bottles of platelet dry powder, each bottle containing about 3 billion platelets .

A bottle of platelet dry powder was taken, and immediately rehydrated according to the method described in Example 1, and the number of platelets was measured. The results are shown in Table 6. The remaining bottles were immediately vacuum-packed and sub-sterilized.

Preliminary Example 17:

The procedure was the same as in Preparation Example 16, except that 30 mL of a physiological saline solution containing 1 mM of ADP was used instead of 30 mL of a reverse osmosis aqueous solution containing 1 mM of ADP.

The number of platelets was determined in the same manner as in Example 16 and the test results are shown in Table 6.

Preliminary Example 18:

The procedure is the same as in Example 16, but 30 mL of ADP containing 1 mM The saline solution was changed to a mixed solution of 15 mL of a physiological saline solution containing 1 mM of ADP and 15 mL of a reverse osmosis aqueous solution containing 1 mM of ADP.

The number of platelets was determined in the same manner as in Example 16 and the test results are shown in Table 6.

Preliminary Example 19:

The procedure was the same as in Example 16, except that the PLT of the plasma layer was centrifuged and removed, and diluted to 20 ml with physiological saline.

The number of platelets was determined in the same manner as in Example 16 and the test results are shown in Table 6.

Preliminary Example 20:

The procedure was the same as in Example 16, except that the PLT of the plasma layer was centrifuged and removed, and 10 mL of physiological saline was added, and then diluted with reverse osmosis water to 20 ml.

The number of platelets was determined in the same manner as in Example 16 and the test results are shown in Table 6.

Preliminary Example 21:

The procedure was the same as in Example 17, except that the PLT of the plasma layer was centrifuged and removed, and diluted to 20 ml with physiological saline.

The number of platelets was determined in the same manner as in Example 16 and the test results are shown in Table 6.

Preliminary Example 22:

The procedure was the same as in Example 17, except that the PLT of the plasma layer was centrifuged and removed, and 10 mL of physiological saline was added, and then diluted with reverse osmosis water to 20 ml.

The number of platelets was determined in the same manner as in Example 16 and the test results are shown in Table 6.

Preliminary Example 23:

The procedure was the same as in Example 18, except that the PLT of the plasma layer was centrifuged and removed, and diluted to 20 ml with physiological saline.

The number of platelets was determined in the same manner as in Example 16 and the test results are shown in Table 6.

Preliminary Example 24:

The procedure was the same as in Example 18, except that the PLT of the plasma layer was centrifuged and removed, and 10 mL of physiological saline was added, and then diluted with reverse osmosis water to 20 ml.

The number of platelets was determined in the same manner as in Example 16 and the test results are shown in Table 6.

Examples 16-24:

The dry powder complex of the preliminary examples 16-24 was prepared with 4 mL of reverse osmosis water, respectively. Water, forming a complex aqueous solution 16~24.

HA blocks 16~24 (weight as shown in Table 8) were immersed in the rehydration solution 16~24, vacuumed for about 20 minutes, filtered, and then sent to the -40 °C freezer, frozen for 24 hours, taken out and weighed. The weight is shown in Table 8. Then, it was sent to a freeze dryer for freeze-drying for 36 hours to obtain HA blocks 16 to 24 coated with freeze-dried platelets.

On the day of production, soak the HA block 16~24 with 10 mL of reverse osmosis water, vacuum and continue to soak for about 20 minutes, filter dry; repeat the soaking and drying steps 6 times; respectively, 7 times of the obtained soaking solution Concentrate and add water to dilute to 80 mL (extracted solution 16~24).

According to the method described in Example 1, the number of platelets of the extracts 16 to 24 was measured. The results are shown in Table 6. The normalization formula is as follows. The results are shown in Table 6: N = N _j * 80 / W _j

Wherein N is the normalized PDGF titer; N _j represents the measured value of the PDGF titer of the HA block, see Table 6; W _j is the weight of the platelet solution soaked in the HA block, see Table 6; 80 is because 80 mL The weight is 80 g.

The experimental data showed that the number of platelets after normalization of HA blocks 16 to 24 was quite consistent with the number of platelets in the preliminary examples 16 to 24, and the ratios were 0.971, 0.963, 0.981, 0.952, 0.984, 0.961, 0.974, and 0.964, respectively. 0.947.

Claims (18)

A method for coating freeze-dried platelets on an implant, the method of coating the protected freeze-dried platelets on the implant, the coating method comprising: a protection step of adding a protective agent to the blood or blood preparation; and separating the plasma a step of centrifuging the blood or blood preparation to which the protective agent is added, and removing the plasma layer to form a platelet thick liquid; a dilution step, which is in a platelet thick liquid, adding an appropriate amount of water or an aqueous solution to form a protected a platelet solution; a coating step of coating the protected platelet solution on the surface of the implant; and a lyophilization step of lyophilizing the platelet-coated solution to freeze the platelet solution coated on the surface of the implant Dry into freeze-dried platelets. The method of claim 1, wherein the protected platelet solution is further lyophilized to form a platelet dry powder, and then an appropriate amount of water or an aqueous solution is added to form a protected platelet solution. The method of claim 1 or 2, wherein the protected platelet solution has a platelet concentration of 50 K to 20 M/μL. The method of claim 3, wherein the protected platelet solution has a platelet concentration of 100 K to 10 M/μL. The method of claim 4, wherein the protected platelet solution has a platelet concentration of 200 K to 5 M/μL. The method of claim 1, wherein the protected platelet solution is protected with adenosine diphosphate. The method of claim 1 or 2, wherein the protected platelet solution is a platelet solution prepared by an ADP protection program, wherein the ADP protection program produces a platelet thick solution: in a mammal Animal platelet thick plasma is added with 0.1 to 10 mM ADP solution and stirred evenly to protect platelets. The ADP dosage is 0.05 to 5 Nemo ADP per plate of platelet-rich plasma in platelet-rich plasma; The protected platelet-rich plasma is removed from the plasma layer to obtain a protected platelet thick solution. The method of claim 7, wherein the ADP solution has a concentration of 0.2 to 5 mM, and the ADP is used in an amount of 0.1 to 2 nanomolar ADP per 1 million platelet cells in the platelet-rich plasma. The method of claim 8, wherein the ADP solution has a concentration of 0.5 to 2 mM, and the ADP is used in an amount of 0.2 to 1 nanomolar ADP per 1 million platelet cells in the platelet-rich plasma. The method of claim 1 or 2, wherein the covering step is by dipping coating, spraying or brushing. The method of claim 1 or 2, wherein the platelets are autologous platelets, allogeneic platelets, or heterogeneous platelets taken from cattle, pigs or sheep. An implant coated with freeze-dried platelets, the body of which is a medical implant characterized in that the surface of the implant is coated with a layer of protected freeze-dried platelets. The implant according to claim 12, wherein the implant has a freeze-dried platelet amount of 20 M to 10000 M per square centimeter of the implant to be coated. The implant according to claim 13, wherein the freeze-dried platelet contained in the implant is coated with 100 M to 5000 M platelets per square centimeter of the area to be covered of the implant. The implant of claim 14, wherein the platelet is autologous platelets, allogeneic platelets, or heterogeneous platelets taken from cattle, pigs or sheep. The implant of claim 12, wherein the protected freeze-dried platelets are microcoagulated with an ADP solution and then lyophilized. The implant of claim 12, wherein the protected freeze-dried platelet layer is made by coating a protected platelet thick solution or a diluted solution thereof, or by reconstituting the protected platelet dry powder. Platelet solution, formed by lyophilization. The implant of claim 17, wherein the protected freeze-dried platelet layer is made by coating a protected platelet thick solution or a diluted solution thereof, or by reconstituting the protected platelet dry powder. The platelet solution is prepared by an ADP protection program; wherein the ADP protection program produces a platelet thick solution by adding 0.1 to 10 mM ADP solution to the thick plasma of mammalian platelets and stirring evenly to protect the platelets. Wherein the amount of ADP is 0.05 to 5 nanomolar ADP per 1 million platelet cells in platelet-rich plasma; and the protected platelet-rich plasma is removed from the plasma layer to obtain a protected platelet thick solution.