Platelet-rich plasma (PRP): methodological aspects and clinical applications

The clinical use of platelet-rich plasma (PRP) is based on the increase in the concentration of growth factors and in the secretion of proteins which are able to maximize the healing process at the cellular level. Since PRP is an autologous biologic material, it involves a minimum risk of immune reactions and transmission of infectious and contagious diseases, and it has been widely used for the recovery of musculoskeletal lesions. Despite the great potential for applicability, the implementation of the therapeutic employment of PRP as a clinical alternative has become difficult, due to the lack of studies related to the standardization of the techniques and/or insufficient description of the adopted procedures. Therefore, it is required establish standard criteria to be followed for obtaining a PRP of high quality, as well as a larger number of studies which should establish the proper concentration of platelets for the different clinical conditions. In this context, the purpose of this review is to discuss some methodological aspects used for achieving the PRP, as well as to discuss the bioactive properties of PRP, and to point out its therapeutic use in different fields of regenerative medicine.

Chitosan-alginate membranes accelerate wound healing

The purpose of this study was to evaluate the efficacy of chitosan-alginate membrane to accelerate wound healing in experimental cutaneous wounds. Two wounds were performed in Wistar rats by punching (1.5 cm diameter), treated with membranes moistened with saline solution (CAM group) or with saline only (SL group). After 2, 7, 14, and 21 days of surgery, five rats of each group were euthanized and reepithelialization was evaluated. The wounds/scars were harvested for histological, flow cytometry, neutrophil infiltrate, and hydroxyproline analysis. CAM group presented higher inflammatory cells recruitment as compared to SL group on 2nd day. On the 7th day, CAM group showed higher CD11b+ level and lower of neutrophils than SL group. The CAM group presented higher CD4+ cells influx than SL group on 2nd day, but it decreased during the follow up and became lower on 14th and 21st days. Higher fibroplasia was noticed on days 7 and 14 as well as higher collagenesis on 21st in the CAM group in comparison to SL group. CAM group showed faster reepithelialization on 7th day than SL group, although similar in other days. In conclusion, chitosan-alginate membrane modulated the inflammatory phase, stimulated fibroplasia and collagenesis, accelerating wound healing process in rats.

New LLLT protocol to speed up the bone healing process-histometric and immunohistochemical analysis in rat calvarial bone defect

Conselho Nacional de Desenvolvimento Científico e Tecnológico (CNPq)

Effects of oestrogen deficiency and 17 beta-estradiol therapy on bone healing in calvarial critical size defects treated with bovine bone graft

Fundação de Amparo à Pesquisa do Estado de São Paulo (FAPESP)

Raloxifene enhances peri-implant bone healing in osteoporotic rats

Fundação de Amparo à Pesquisa do Estado de São Paulo (FAPESP)

Hypertension modifies OPG, RANK, and RANKL expression during the dental socket bone healing process in spontaneously hypertensive rats

Fundação de Amparo à Pesquisa do Estado de São Paulo (FAPESP)

Diabetes reduces mesenchymal stem cells in fracture healing through a TNF alpha-mediated mechanism

Diabetes interferes with bone formation and impairs fracture healing, an important complication in humans and animal models. The aim of this study was to examine the impact of diabetes on mesenchymal stem cells (MSCs) during fracture repair.Fracture of the long bones was induced in a streptozotocin-induced type 1 diabetic mouse model with or without insulin or a specific TNF alpha inhibitor, pegsunercept. MSCs were detected with cluster designation-271 (also known as p75 neurotrophin receptor) or stem cell antigen-1 (Sca-1) antibodies in areas of new endochondral bone formation in the calluses. MSC apoptosis was measured by TUNEL assay and proliferation was measured by Ki67 antibody. In vitro apoptosis and proliferation were examined in C3H10T1/2 and human-bone-marrow-derived MSCs following transfection with FOXO1 small interfering (si)RNA.Diabetes significantly increased TNF alpha levels and reduced MSC numbers in new bone area. MSC numbers were restored to normal levels with insulin or pegsunercept treatment. Inhibition of TNF alpha significantly reduced MSC loss by increasing MSC proliferation and decreasing MSC apoptosis in diabetic animals, but had no effect on MSCs in normoglycaemic animals. In vitro experiments established that TNF alpha alone was sufficient to induce apoptosis and inhibit proliferation of MSCs. Furthermore, silencing forkhead box protein O1 (FOXO1) prevented TNF alpha-induced MSC apoptosis and reduced proliferation by regulating apoptotic and cell cycle genes.Diabetes-enhanced TNF alpha significantly reduced MSC numbers in new bone areas during fracture healing. Mechanistically, diabetes-enhanced TNF alpha reduced MSC proliferation and increased MSC apoptosis. Reducing the activity of TNF alpha in vivo may help to preserve endogenous MSCs and maximise regenerative potential in diabetic patients.

Healing at implant sites prepared conventionally or by means of Sonosurgery (R): an experimental study in dogs

ObjectiveTo compare peri-implant tissue healing at implants installed in sites prepared with conventional drills or a sonic device.Material and methodsIn six Beagle dogs, the mandibular premolars and first molars were extracted bilaterally. After 3 months, full-thickness muco-periosteal flaps were elevated and recipient sites were prepared in both sides of the mandible. In the right side (control), the osteotomies were prepared using conventional drills, while, at the left side (test), a sonic device (Sonosurgery((R))) was used. Two implants were installed in each side of the mandible. After 8weeks of non-submerged healing, biopsies were harvested and ground sections prepared for histological evaluation.ResultsThe time consumed for the osteotomies at the test was more than double compared to the conventional control sites. No statistically significant differences were found for any of the histological variables evaluated for hard and soft tissue dimensions. Although not statistically significant, slightly higher mineralized bone-to-implant contact was found at the test (65.4%) compared to the control (58.1) sites.ConclusionsSimilar healing characteristics in osseointegration and marginal hard tissue remodeling resulted at implants installed into osteotomies prepared with conventional drills or with the sonic instrument (Sonosurgery((R))).

Healing of implants installed in over- or under-prepared sites: an experimental study in dogs

ObjectiveTo study bone healing at implants installed with different insertion torques.Material and methodsIn six Labrador dogs, all mandibular premolars and first molars were extracted. After 4months of healing, flaps were elevated, and two implant sites were prepared at each side of the mandible. In the right side of the mandible, the distal sites were prepared conventionally, while the mesial sites were over-prepared by 0.2mm. As a consequence, a final insertion torque of similar to 30Ncm at the distal and a minimal insertion torque close to 0Ncm at the mesial sites were obtained. In the left sides of the mandible, however, the recipient sites were underprepared by 0.3mm resulting in an insertion torque of 70Ncm at both implants. Cover screws were applied, and flaps sutured to fully submerge the experimental sites. After 4months, the animals were sacrificed and ground sections obtained for histological evaluation.ResultsThe mineralized bone-to-implant contact was in the range of 55.2-62.1%, displaying the highest value at implants with similar to 30Ncm insertion torque and the lowest value at the implant sites with close to 0Ncm insertion torque. No statistically significant differences were revealed. Bone density was in the range of 43.4-54.9%, yielding the highest value at implants with 70Ncm insertion torque and the lowest at the implant sites with close to 0Ncm insertion torque. The difference between the sites of similar to 30Ncm and the corresponding 70Ncm insertion torque reached statistical significance.ConclusionsSimilar amounts of osseointegration were obtained irrespective of the insertion torque applied. Moreover, implants installed in sites with close to 0Ncm insertion torque may properly osseointegrate as well.

Sequential healing of the elevated sinus floor after applying autologous bone grafting: an experimental study in minipigs

AimTo describe the sequential healing after elevation of the maxillary sinus mucosa applying the lateral access technique with the use of autogenous bone grafting without membrane to occlude the osteotomy access.Material and methodsImmediately after the elevation of the maxillary sinus Schneiderian membrane, applying the lateral access technique in 10 minipigs, autologous bone was harvested from the lateral aspect of the mandibular molar region and ground into particles with a bone mill. The space under the Schneiderian membrane was filled with this graft. No membranes were placed onto the access osteotomy. The healing was evaluated after 15, 30, 90 and 180days. Paraffin sections were prepared and analyzed histologically.ResultsAfter 15days of healing, the elevated area was mainly filled with provisional matrix, newly formed bone and some remnants of bone chips, and appeared reduced in volume compared with that at the time of surgery. After 30days of healing, further shrinkage of the height of the elevated space was found, with similar percentages of the different tissue components. After 90 and 180days, the area underneath the Schneiderian membrane appeared reduced in volume and condensed toward the base of the sinus. The bone tissues appeared to be more mature, both for the mineralized and the non-mineralized portions, while connective tissue occupied 20% of the space, most likely related to the lack of the use of a membrane occluding the access at the time of surgery.ConclusionsSuboptimal healing outcomes with respect to augmentation of the space under the sinus floor membrane were documented when autologous bone chips were used as a filler and no membrane was applied to cover the access.

Healing at mandibular block-grafted sites: an experimental study in dogs

AimThe aim of this study was to evaluate the healing of autologous bone block grafts or deproteinized bovine bone mineral (DBBM) block grafts applied concomitantly with collagen membranes for horizontal alveolar ridge augmentation.Material and methodsIn six Labrador dogs, molars were extracted bilaterally, the buccal bony wall was removed, and a buccal box-shaped defect created. After 3months, a bony block graft was harvested from the right ascending ramus of the mandible and reduced to a standardized size. A DBBM block was tailored to similar dimensions. The two blocks were secured with screws onto the buccal wall of the defects in the right and left sides of the mandible, respectively. Resorbable membranes were applied at both sides, and the flaps sutured. After 3months, one implant was installed in each side of the mandible, in the interface between grafts and parent bone. After 3months, biopsies were harvested and ground sections prepared to reveal a 6-month healing period of the grafts.Results776.2% and 5.9 +/- 7.5% of vital mineralized bone were found at the autologous bone and DBBM block graft sites, respectively. Moreover, at the DBBM site, 63 +/- 11.7% of connective tissue and 31 +/- 15.5% of DBBM occupied the area analyzed. Only 0.2 +/- 0.4% of DBBM was found in contact with newly formed bone. The horizontal loss was in a mean range of 0.9-1.8mm, and 0.3-0.8mm, at the autologous bone and DBBM block graft sites, respectively.ConclusionsAutologous bone grafts were vital and integrated to the parent bone after 6months of healing. In contrast, DBBM grafts were embedded into connective tissue, and only a limited amount of bone was found inside the scaffold of the biomaterial.

Chitosan-alginate membranes accelerate wound healing

Coordenação de Aperfeiçoamento de Pessoal de Nível Superior (CAPES)

Radiographic characterization of the bone healing process after protein application extracted from hevea brasiliensis

Fundação de Amparo à Pesquisa do Estado de São Paulo (FAPESP)

Healing process of dogs' pulpless teeth after apicoectomy and root canal filling at different levels

Dogs' teeth with infected root canals, were submitted to apicoectomy and the root canals were filled flush or 2 mm short. In a third group the canals were left unfilled and only the access openings were closed. One hundred and eighty days after the treatment, the animals were killed and the specimens prepared for histological analysis. Repair was not observed in the teeth with unfilled root canals. The healing observed in the teeth with root canals filled flush was less complete than the healing observed when the root canals were filled 2 mm short.

Effect of root canal status on periodontal healing after surgical injury in dogs.

This study was carried out to observe if the status of the root canal might influence the healing process of surgically prepared experimental periodontal lesions. Forty tooth roots from four dogs were divided into four different groups: a) root canals with vital pulps, b) root canals open to the oral environment, c) root canals infected and filled with zinc oxide eugenol cement, and d) root canals infected and filled with calcium hydroxide. By means of a surgical intervention, a cavity was prepared in the medium portion of the roots. Six months later, the specimens were removed and prepared for histological analysis. The results, which were submitted to statistical analysis, showed that the status of the root canals influenced the healing process of the experimental periodontal lesions. In the groups where the root canals were filled, calcium hydroxide gave the best results. In the group with root canals left open to the oral environment, resorption of the dentin of the experimental cavities, was the most obvious observation. However, it did not prevent the repair process, only slowed it down.