Autogenous Platelet-rich Plasma Research Articles

Therapeutic Effect of Equine Musculoskeletal Disease using Autogenous Platelet Rich Plasma

This study evaluated the therapeutic effect using autogenous platelet-rich plasma (PRP) for the musculoskeletal disease of Thoroughbred horses raised in South Korea. Twenty-five Thoroughbred horses (15 adult horses more than 2 years old, 10 fostering horses less than 2 years old) were diagnosed with musculoskeletal disorders (2 horses with desmitis of the suspensory ligament, 9 horses with osteoarthritis, 14 horses with fracture) by ultrasound and X-ray were used in this study. We collected venous blood of patients by using ACP Double Syringe after sterilization of jugular site, and then autogenous PRP was taken through centrifugation. We were observed the healing process of the lesion by using ultrasonography and X-ray images after injecting 4mL of autogenous PRP into each lesion for 2-times. Results of this study, 20 (80%) of 25 damaged horses returned to normal competition. The core lesion in the ligament disappeared in 90 days and healthy tissue regenerated. The fracture line of the cortex of bone disappeared in 60 days, and no side effects such as osteophytosis during healing progress. Remodeled articular surface, and cartilage defect, a fragment in joint disappeared. It could be confirmed that applying autogenous PRP treatment is an excellent regenerative therapy for equine locomotorium lesions.

Clinical Outcome and 8-Year Follow-Up of Alveolar Bone Tissue Engineering for Severely Atrophic Alveolar Bone Using Autologous Bone Marrow Stromal Cells with Platelet-Rich Plasma and β-Tricalcium Phosphate Granules.

Background: Although bone tissue engineering for dentistry has been studied for many years, the clinical outcome for severe cases has not been established. Furthermore, there are limited numbers of studies that include long-term follow-up. In this study, the safety and efficacy of bone tissue engineering for patients with a severely atrophic alveolar bone were examined using autogenous bone marrow stromal cells (BMSCs), and the long-term stability was also evaluated. Methods: BMSCs from iliac bone marrow aspirate were cultured and expanded. Then, induced osteogenic cells were transplanted with autogenous platelet-rich plasma (PRP) and β-tricalcium phosphate granules (β-TCP) for maxillary sinus floor and alveolar ridge augmentation. Eight patients (two males and six females) with an average age of 54.2 years underwent cell transplantation. Safety was assessed by monitoring adverse events. Radiographic evaluation and bone biopsies were performed to evaluate the regenerated bone. Results: The major population of transplanted BMSCs belonged to the fraction of CD34−, CD45dim, and CD73+ cells, which was only 0.065% of the total bone marrow cells. Significant deviations were observed in cell growth and alkaline phosphatase activities among individuals. However, bone regeneration was observed in all patients and the average bone area in the biopsy samples was 41.9% 6 months following transplantation, although there were also significant deviations among each case. No adverse events related to the transplants were observed. In the regenerated bone, 27 out of 29 dental implants were integrated. Dental implants and regenerated bone were stable for an average follow-up period of 7 years and 10 months. Conclusions: Although individual variations were observed, the results showed that bone tissue engineering using BMSCs with PRP and β-TCP was feasible for patients with severe atrophic maxilla throughout a long-term follow-up period and was considered safe. However, further studies with a larger number of cases and controls to confirm the efficacy of BMSCs and the development of a protocol to establish a reproducible quality of stem cell-based graft material will be required.

Use of autogenous platelet rich plasma and 3D collagen matrix Fibromatrix for soft tissue regeneration in the treatment of Miller Class I gingival recessions

Background: Evaluation of the effectiveness of the integrated use of autogenous platelet plasma and 3D collagen matrix Fibromatrix for the regeneration of soft tissues of the oral cavity in the surgical treatment of Miller Class I gingival recession. Materials and Method: 25 patients were examined and treated. All patients underwent surgical treatment of gingival recession using two-layer techniques of eliminating gingival recession with «3D collagen matrix FibroMATRIX» for the regeneration of soft tissues of the oral cavity and autologous platelet plasma. In all cases, the rehydration of the collagen matrix was carried out in autogenous platelet plasma for 2 minutes. The result was evaluated after 7th day, 14th day, 1 and 3 months. Statistical processing of the studies was carried out in GraphPadPrism v.6.0, using the t-test, Mann-Whitney. Differences were considered significant at p <0.05. Results: Assessment of gingival recession closure and its significant (p <0.05) increase, corresponding to 85% efficiency after treatment in both groups of patients, no significant differences were found in both groups. The best clinical results were achieved in eliminating gingival recessions in the anterior maxilla, showing its 100% efficiency. Conclusions: The combined use of autogenous platelet plasma and 3D collagen matrix Fibromatrix for the regeneration of soft tissues of the oral cavity is an effective method for treating Miller Class I gingival recession.

Comparative evaluation of efficacy of autogenous platelet rich plasma versus visco supplementation in treatment of early osteoarthritis of knee

<p class="abstract"><strong>Background:</strong> Osteoarthritis is a very common chronic degenerative disease most commonly affecting the knee joints. In present study we compared the efficacy of autogenous platelet rich plasma (PRP) versus visco supplementation in treatment of early osteoarthritis of knees.</p><p class="abstract"><strong>Methods:</strong> 30 patients (56 knees) were registered and divided into two groups. Out of which PRP in 28 knees and visco supplementation in 28 knees injected during. Visual analogue scale (VAS) and Western Ontario and McMaster Universities Arthritis Index (WOMAC) scores were measured. These scores were measured at first visit, 6 weeks, 12 weeks and at 24 weeks.<strong></strong></p><p class="abstract"><strong>Results:</strong> All registered patients were randomized in two groups. Group I (total 16 patients and 28 knees) for intraarticular PRP injection and group II (total 14 patients and 28 knees) for intraarticular viscosupplement injection. Out of 28 knees of group I; 12 (42.85%) knees belonged to grade II and 16 (57.15%) of grade III. Out of 28 knees of group II; 15 (53.57%) knees of grade II and 13 (46.45%) of grade III. None of the knees belonged to grade I and grade 0. There was significant difference in outcome of two treatment groups (p<0.05) at 24 weeks.</p><p class="abstract"><strong>Conclusions:</strong> Treatment with PRP showed a significantly better clinical outcome compared with viscosupplemention at 24 weeks follow up. Although patients achieved lower WOMAC and VAS scores in PRP group at 6 and 12 weeks follow up that was statistically insignificant. We conclude that long term results of PRP are better than viscosupplementation.</p>

Platelet-rich plasma promotes the regeneration of cartilage engineered by mesenchymal stem cells and collagen hydrogel via the TGF-β/SMAD signaling pathway.

The tissue engineering technique using mesenchymal stem cells (MSCs) and scaffolds is promising. Transforming growth factor-β1 (TGF-β1) is generally accepted as an chondrogenic agent, but immunorejection and unexpected side effects, such as tumorigenesis and heterogeneity, limit its clinical application. Autogenous platelet-rich plasma (PRP), marked by low immunogenicity, easy accessibility, and low-cost, may be favorable for cartilage regeneration. In our study, the effect of PRP on engineered cartilage constructed by MSCs and collagen hydrogel in vitro and in vivo was investigated and compared with TGF-β1. The results showed that PRP promoted cell proliferation and gene and protein expressions of chondrogenic markers via the TGF-β/SMAD signaling pathway. Meanwhile, it suppressed the expression of collagen type I, a marker of fibrocartilage. Furthermore, PRP accelerated cartilage regeneration on defects with engineered cartilage, advantageous over TGF-β1, as evaluated by histological analysis and immunohistochemical staining. Our work demonstrates that autogenous PRP may substitute TGF-β1 as a potent and reliable chondrogenic inducer for therapy of cartilage defect.

Effect of Platelet-rich Plasma on Implant Stability in the Mandible

Background and aims. Plasma rich in growth factors (PRGFs) has been recently proposed as an aid to enhance regeneration of osseous and epithelial tissues in oral surgery. The purpose of this study was to determine the effect of local application of platelet-rich plasma (PRP) on implant stability measured by periotest. Materials and methods. A total of 24 implants were placed in the mandibles of 12 lower edentulous patients. In each patient, 2 implants were placed anterior to the mental foramen in bilateral canine sites. One implant in each patient was dipped in autogenous PRP before insertion (test group), while the other implant was not embedded in PRP (control group). Repeated stability measurements were carried out by periotest on the day of surgery and 1, 2, 4 and 8 weeks after surgery. Results. In both groups minimum periotest values (highest stability) were observed on the day of surgery and 8 weeks after surgery. The maximum periotest values (lowest stability) were observed in 4th week after surgery. Considering implant stability, no statistically significant differences were observed between the test and control groups at any time (P>0.05). In the PRP group, the difference in implant stability between the day of surgery and the 2nd and 4th weeks were statistically significant (P<0.05). Conclusion. Application of PRP on implant surface did not have any additional effect on implant stability in the mandible

The Role of Platelet Rich Plasma on Healing of Induced Femoral Fracture Fixed With Dynamic Compression Plate in Dogs.

The present study was carried out to evaluate the platelet rich plasma (PRP) as a factor for accelerating bone healing and compared with normal healing in 18 apparently healthy male mongrel dogs based on clinical, radiographic and histopathological findings. Induction of femoral fracture was done in all dogs of both groups, a gap (0.5 cm × 0.5cm with the whole depth of the near cortex) was created at the fracture sit bone rongeur and the fracture was fixed with standard dynamic compression plate (DCP) and self taping cortical screws. Dogs were divided into two equal groups, each of 9 dogs. Group I was left as a control. In group II, the gap was packed with autogenous PRP. The results of this study pointed out that PRP slightly improved bone healing.

Comparison of Healing of Bone Defects by Autogenous Platelet Rich Plasma with Hydroxyapatite and Natural Healing in Pediatric Patients

Comparison of Healing of Bone Defects by Autogenous Platelet Rich Plasma with Hydroxyapatite and Natural Healing in Pediatric Patients

Adipose-derived stem cells incorporated into platelet-rich plasma improved bone regeneration and maturation in vivo.

Some cases of tooth loss related to dental trauma require bone-grafting procedures to improve the aesthetics before prosthetic rehabilitation or to enable the installation of dental implants. Bone regeneration is often a challenge and could be largely improved by mesenchymal stem cells therapy. However, the appropriate scaffold for these cells still a problem. This study evaluated the in vivo effect of human adipose-derived stem cells incorporated into autogenous platelet-rich plasma in bone regeneration and maturation. Adipose-derived stem cells were isolated from lipoaspirate tissues and used at passage 4. Immunophenotyping and multilineage differentiation of cells were performed and mesenchymal stem cells characteristics confirmed. Bicortical bone defects (10 mm diameter) were created in the tibia of six beagle dogs to evaluate the effect of adipose-derived stem cells incorporated into platelet-rich plasma scaffolds, platelet-rich plasma alone, autogenous bone grafts, and clot. Samples were removed 6 weeks postsurgeries and analyzed by quantification of primary and secondary bone formation and granulation tissue. Adipose-derived stem cells incorporated into platelet-rich plasma scaffolds promoted the highest bone formation (primary + secondary bone) (P < 0.001), the highest bone maturation (secondary bone) (P < 0.001), and the lowest amount of granulation tissue (P < 0.001). Adipose-derived stem cells incorporated into platelet-rich plasma scaffolds promote more bone formation and maturation, and less granulation tissue in bone defects created in canine tibia. Therefore, platelet-rich plasma can be considered as a candidate scaffold for adipose-derived stem cells to promote bone regeneration.

Autogenous Platelet Rich Plasma in Healing of Bone Defects

Purpose: To evaluate the efficacy of PRP in regeneration of bone in defects of the oral and maxillofacial region.&#x0D; Methods: A comparative nonrandomized study including 40 patients with bony defects of cystic origin in the maxillofacial region was carried out in the Department of Oral and Maxillofacial Surgery. Patients were divided into two groups: group A received PRP and in group B PRP was not used. Postoperatively, the patients were monitored regularly by radiographs to evaluate new bone growth at 1st ,3rd and 6th month clinically and radiographically.&#x0D; Results: Faster bone healing in was observed in the patients in whom PRP was used at every follow up.&#x0D; Conclusion: Defects filled with PRP showed comparatively earlier and faster bone regeneration. JMS 2013;16(2):90-94

Allografts with Autogenous Platelet-Rich Plasma for Tibial Defect Reconstruction: A Rabbit Study

To evaluate the effect of autogenous platelet-rich plasma (PRP) for fresh-frozen allografts in tibial defect reconstruction in rabbits. 40 adult New Zealand white rabbits underwent tibial defect reconstruction with autografts (n=12), allografts without PRP (n=12), or allografts with PRP (n=12) and were observed for 12, 16, and 24 weeks (4 for each period). Tibias of the remaining 4 rabbits were used as donor allografts, and the remaining allografts were procured from recipient rabbits. A 1.5- cm cortical segment of the tibia was osteotomised, and then fixed with a 9-hole mini-compression plate and 2 cerclage wires. Allografts were stripped off the periosteum and soft tissues and medullary contents, and then stored in a freezer at -80 ºC. All allografts were deep frozen for at least 4 weeks before transplantation. 7 ml of whole blood was drawn to prepare 1 ml of PRP. The PRP was then mixed with 1.0 ml of human thrombin to form a platelet gel. The PRP gel was then packed into the medullary canal of the allograft and applied on the cortical surface before tibial defect reconstruction. Rabbits were sacrificed at 12, 16, and 24 weeks. The specimens were assessed for bone union at host-graft junctions and for bone resorption, new bone formation, callus encasement, and viable osteocyte counts. There were 4 specimens in each group at each observation period. Osteoid bridging the gap at host-graft junctions was noted in all specimens in the autograft and allograft-with-PRP groups at week 12 and in the allograft-without-PRP group at week 24. Bone union in allografts without PRP was delayed. All indices for biological incorporation (resorption index, new bone formation index, callus encasement index, and viable osteocyte count) were significantly greater in the autograft than allograft-without-PRP groups, except for the resorption index at week 24, whereas the differences were not significant between the autograft and allograft-with-PRP groups. The differences between the 2 allograft groups were usually not significant, except for the resorption index. PRP-augmented allografts behaved similarly to autografts for tibial defect reconstruction in rabbits. PRP increased bone union and bone resorption.

Torus mandibularis bone chips combined with platelet rich plasma gel for treatment of intrabony osseous defects: clinical and radiographic evaluation

The use of platelet rich plasma (PRP) gel in combination with torus mandibularis offers a potentially useful treatment for periodontal osseous defects. Whether this combination enhances the outcome of periodontal regenerative therapy is not known. This study compared the effectiveness of torus mandibularis bone chips alone and when combined with autogenous PRP gel in treating periodontal osseous defects. 24 sites from 12 patients were selected using a split mouth design and determined by a double-blind, randomized, controlled clinical trial. Both sites received a full-thickness mucoperiosteal flap; one intrabony defect was filled with torus mandibularis bone chips alone and the other with torus mandibularis bone chips mixed with PRP gel. There was a 57% gain in the clinical attachment level and 60% reduction in the probing depth for torus mandibularis alone compared to 72% and 68% for sites treated with torus mandibularis and PRP gel (p≤0.01). There was a statistically significant difference in the bone dentistry and the marginal bone loss at sites with PRP gel compared to those without gel (p≤0.01). The use of mandibular tori as autogenous bone graft combined with PRP gel showed a significant improvement in the clinical outcome of periodontal therapy than mandibular tori alone.

Modificação de técnica de preparo do plasma rico em plaquetas em cães

Platelet-rich plasma (PRP) has many growing factors that improve tissue regeneration. The goal of this paper is to describe an innovative, quick and simple technique for preparing autogenous PRP from a small amount of blood, which allowing use in dogs. After harvesting 8ml of total blood in five dogs, PRP was obtained using an aseptic technique, which rendered a platelet count two to six times more concentrated than the previous serum platelet count, with final absolute values between 800.000 and 1.600.000/mm3. The technique can be developed quickly, without complications using simple equipments, reaching an ideal platelet concentration and keeping plasma sterility.

Healing of Fresh Frozen Bone Allograft with or without Platelet‐Rich Plasma: A Histologic and Histometric Study in Rats

This study histomorphometrically analyzed the effect of autogenous platelet-rich plasma (PRP) on healing of fresh frozen bone allograft (FFBA) in bony defects in rat calvaria. A 5mm-diameter defect was created in the calvarium of 30 rats. Animals were divided into three groups: C (defect was filled by blood clot only), FFBA (defect was filled with 0.01mL of FFBA), and FFBA/PRP (defect was filled with 0.01mL of FFBA combined with 100µL of PRP). All animals were euthanized at 30 days postoperatively. Histomorphometry and histology analyses were performed. Data were statistically analyzed (analysis of variance, Tukey, p<.05). FFBA had a statistically smaller new bone area than groups FFBA/PRP and C. No statistically significant differences were observed between groups FFBA and FFBA/PRP with regard to remaining bone graft particle area. It can be concluded that (1) PRP improved the incorporation of FFBA, increasing the amount of new bone formed; (2) PRP has not influenced the resorption of nonviable particles of the FFBA; and (3) presence of remaining FFBA particles might have accounted for the smaller amount of new bone observed in group FFBA when compared with control group.

Increased survival of free fat grafts with platelet-rich plasma in rabbits

Numerous applications of autogenous platelet-rich plasma (PRP) have been studied so far; however, its property of enhancing the survival of free fat grafts has not been defined yet. In the literature, many reports are anecdotal and few include controls to definitely determine the role played by PRP in these grafts. PRP was investigated to study its effect in free fat grafts' survival in a rabbit model. A total of 30 New Zealand male rabbits aged 6 months received 0.8 g fat tissue (harvested by scissors dissection from the scapular area of the own animal) in the ears and were randomised into two groups. Group 1 (PRP group) was given the combination of 0.8 g of free fat graft and 1 ml of PRP. Group 2 (control group) received 0.8 g of free fat graft and 1 ml of saline solution. The rabbits were followed up for a period of 6 months after the procedure and then euthanised. The grafted tissue was stained with haematoxylin-eosin and submitted to microscopical evaluation. Graft histopathology was investigated for adipocyte viability, number of blood vessels and the presence of necrosis and fibrosis. All data were statistically analysed by the differences between the study groups. Three major effects of the addition of PRP in the free fat graft were observed. Group 1 showed a significantly higher fat survival weight as compared with the control group (P<0.05). Histopathological investigations revealed that the number of viable adipocytes and blood vessels was higher in group 1, and still, a larger number of necrotic areas and fibrosis were detected between group 2 (P<0.05). Application of autogenous PRP can enhance free fat graft survival in rabbits.

Effect of platelet‐rich plasma on bone healing of autogenous bone grafts in critical‐size defects

This study histologically analysed the effect of autogenous platelet-rich plasma (PRP), prepared according to a new semiautomatic system, on healing of autogenous bone (AB) grafts placed in surgically created critical-size defects (CSD) in rabbit calvaria. Sixty rabbits were divided into three groups: C, AB and AB/PRP. A CSD was created in the calvarium of each animal. In Group C (control), the defect was filled by blood clot only. In Group AB (autogenous bone graft), the defect was filled with particulate autogenous bone. In Group AB/PRP (autogenous bone graft with platelet-rich plasma), it was filled with particulate autogenous bone combined with PRP. All groups were divided into subgroups (n=10) and euthanized at 4 or 12 weeks post-operatively. Histometric and histologic analyses were performed. Data were statistically analysed (anova, t-test, p<0.05). Group C presented significantly less bone formation compared with Group AB and AB/PRP in both periods of analysis (p<0.001). At 4 weeks, Group AB/PRP showed a statistically greater amount of bone formation than Group AB (64.44 +/- 15.0%versus 46.88 +/- 14.15%; p=0.0181). At 12 weeks, no statistically significant differences were observed between Groups AB and AB/PRP (75.0 +/- 8.11%versus 77.90 +/- 8.13%; p>0.05). It is notable that the amount of new bone formation in Group AB/PRP at 4 weeks was similar to that of Group AB at 12 weeks (p>0.05). Within its limitation, the present study has indicated that (i) AB and AB/PRP significantly improved bone formation and (ii) a beneficial effect of PRP was limited to an initial healing period of 4 weeks.

Prefabrication of vascularized bone segment with collagen microspheres and autogenous platelet-rich plasma

Prefabrication of vascularized bone segment with collagen microspheres and autogenous platelet-rich plasma

The regenerative effect of platelet-rich plasma on healing in large osteochondral defects

Platelet-rich plasma (PRP), a platelet concentrate made of autogenous blood, has been used to improve bone and soft tissue defect healing in recent years. The aim of this study was to assess the effect of PRP on articular cartilage defects in a rabbit model. Forty-eight osteochondral defects created in the femoropatellar groove were (a) left untreated, (b) treated with autogenous PRP in a poly-lactic-glycolic acid (PLGA), or (c) with PLGA alone. Platelets were enriched 5.12-fold compared to normal blood in the PRP. After four and 12 weeks, the explanted tissue specimens were assessed by macroscopic examination, micro-computed tomography, and histological evaluation. Macroscopic examination, micro-computed tomography and histology of the newly formed cartilage and bone in the defect differ significantly between the PRP-treated and the untreated groups, and stimulatory effect of PRP on osteochondral formation was observed. In conclusion, PRP in PLGA improves osteochondral healing in a rabbit model.

Plasma rico em plaquetas associado ou não ao osso esponjoso autógeno no reparo de falhas ósseas experimentais

O presente estudo avaliou a influência do plasma autógeno rico em plaquetas (PRP), associado ou não ao autoenxerto esponjoso (EOE), na reparação de falhas ósseas criadas no crânio de coelhos. A falha I foi preenchida com o PRP; a falha II com 3mg de EOE; a falha III com EOE associado ao PRP e a falha IV não foi preenchida, servindo como controle. Após as cirurgias, os animais foram separados em três grupos e eutanasiados aos 30, 60 e 90 dias. Na avaliação mesoscópica, independentemente do período de observação, o preenchimento ósseo, na falha controle e naquelas tratadas com PRP, iniciou-se a partir das bordas para o centro e do fundo para a superfície das falhas. Já nas falhas tratadas com EOE e com enxerto associado ao PRP, foi notado também crescimento ósseo na porção central das falhas. Na análise radiográfica, foi observada maior radiopacidade no interior das falhas tratadas com EOE e com enxerto associado ao PRP, em todos os tempos. Microscopicamente, aos 30 dias, na falha tratada com EOE associado ao PRP, os fragmentos ósseos do enxerto estavam indistintos do tecido ósseo neoformado, presente em toda a borda do defeito, associado à moderada quantidade de tecido conjuntivo fibroso muito vascularizado e celularizado. Esse tecido apresentou material amorfo, eosinofílico e extracelular, junto a um processo inflamatório, constituído por linfócitos e, em menor número, por macrófagos e células gigantes multinucleadas, que podem ter influenciado negativamente a formação óssea precoce. Aos 60 e 90 dias, apenas focos ocasionais de inflamação linfocitária foram observados. O comportamento dos dois tratamentos, PRP associado ou não ao EOE, em relação a preenchimento ósseo, foi semelhante ao final do período de observação; o enxerto, utilizado de forma isolada, determinou precocidade de reparação óssea e a tromboplastina, utilizada para formação do gel de plaquetas, incitou uma reação semelhante a do tipo corpo estranho, que atuou negativamente na fase inicial de reparação.

Fabrication of pre-determined shape of bone segment with collagen-hydroxyapatite scaffold and autogenous platelet-rich plasma

Reconstruction of large segment of bony defects is frequently needed in hand surgery. Autogenous bone grafting is considered the standard in management of these bony defects but has limited source of graft material. Collagen and hydroxyapatite have been used as bone-filling materials and are known to serve as the osteoconductive scaffold for bone regeneration. On the other hand, platelet-rich plasma is a kind of natural source of growth factors, and has been used successfully in bone regeneration and improving wound healing. This study was designed to evaluate the effectiveness of using biohybrids of platelet-rich plasma and collagen-hydroxyapatite beads for fabricating of protrusive bone in a rabbit animal model. Biomaterial beads comprised of particulate hydroxyapatite dispersed in fibrous collagen (type I) matrices were prepared and filled in the ringed polytetrafluoroethylene (PTFE) artificial vascular graft (3 cm long, 1 cm in diameter). New Zealand White rabbits were each implanted with two cylindrical PTFE artificial vascular graft over both iliac crests (n = 16). A 2 x 0.5 cm opening was created at the side of each PTFE chamber to allow the content of chamber in contact with the bone marrow of the ileum. The chambers were empty (groups A and D), filled with type I collagen/hydroxyapatite beads (groups B and C). In groups C and D, autologous platelet rich plasma (PRP) was given by transcutaneous injection method into the chambers every week. After 12 weeks, the animals were sacrificed and the chambers were harvested for radiological and histological analysis. In plain radiographs, the group C chambers had significantly higher bone tissue engineered (average calcified density 0.95, average calcified area 61.83%) compared with other groups (P < 0.001). In histological examination, there was a creeping substitution of the implant by the in-growth of fibrovascular tissue in group C. Abundant fibrovascular networks positioned interstitially between these biomaterial beads in all parts of chamber. Degradation of these beads and newly formed capillaries and osteoids around the degraded matrixes are shown. The continually calcification in the matrixes or degraded matrixes is evidenced by the strong green fluorescence observed under the confocal microscope. In group B, looser architecture without evidence of tissue in-growth was shown. In the scaffold absent groups (A and D), there was only fibrous tissue shown within the chamber. In this study, we have demonstrated a feasible approach to fabricate an osseous tissue that meets clinical need. Using the type I collagen/ hydroxyapatite gel beads matrixes and intermittent injection of autologous platelet-rich-plasma, specific 3D osseous tissues with fibrovascular network structure from pre-exist bony margin were successfully created in an in vivo animal model.