Valuation of angiogenesis in bovine xenografts implanted in intracorporal sites of rabbits as models of in-vivo bioreactors / Avaliação da angiogênese em xenoenxertos bovinos implantados em sítios intracorpóreos de coelhos como modelos de biorreatores in vivo

The aim of this study was to evaluate neovascularization of bovine xenografts implanted in intracorporeal sites of rabbits (bioreactors). 30 rabbits were used, divided into 6 groups, according to the evaluation time (7, 15, 30, 45, and 60 days); each animal received xenogenic implants in 3 different intracorporeal sites (A1 - omentum bag; A2 - intermuscular space of quadriceps femoris; A3 - subperiosteal of ilium bone). Histological assessments graded the presence of angiogenesis, the number of inflammatory cells, newly formed bone tissue, and the presence of giant cells. Histological analyses showed intense angiogenesis in all implanted xenografts. Presence of inflammatory infiltrate and giant cells at the A1 implant site and presence of bone neoformation at the A3 implant site were noted. Degeneration of implants and formation of a fibrous capsule were noted. When comparing the interaction of the site with the days of evaluation, statistical analysis showed a significant difference (p≤0.05) in any time of neovascularization analysis. The vascular endothelial growth factor (VEGF) and inflammatory cells of the omentum in its structure, may have contributed to the greater presence of neovessels and inflammatory cells, a fact that may indicate functionality as a possible bone substitute.(AU)

O objetivo deste estudo foi avaliar a neovascularização de xenoenxertos bovinos implantados em sítios intracorpóreos de coelhos (biorreatores). Foram utilizados 30 coelhos, os quais foram divididos em seis grupos, de acordo com o tempo de avaliação (sete, 15, 30, 45 e 60 dias); cada animal recebeu implantes xenogênicos em três diferentes sítios intracorpóreos (A1 - bolsa de omento; A2 - espaço intermuscular do quadríceps femoral; A3 - subperiosteal do osso ílio). Avaliações histológicas classificaram a presença de angiogênese, o número de células inflamatórias, de tecido ósseo neoformado e a presença de células gigantes. As análises histológicas mostraram intensa angiogênese em todos os xenoenxertos implantados. Observou-se presença de infiltrado inflamatório e células gigantes no local do implante A1 e presença de neoformação óssea no local do implante A3. Ao mesmo tempo, a degeneração dos implantes e a formação de uma cápsula fibrosa foram observadas. Ao comparar a interação do local com os dias de avaliação, a análise estatística mostrou diferença significativa (P≤0,05) em qualquer momento da análise de neovascularização. O fator de crescimento endotelial vascular (VEGF) e as células inflamatórias do omento em sua estrutura podem ter contribuído para a maior presença de neovasos e células inflamatórias, fato que pode indicar funcionalidade como possível substituto ósseo.(AU)

Horizontal ridge augmentation of a single atrophic site in the anterior maxilla using hydroxyapatite and rhBMP-2 / Augmento óseo horizontal de reborde alveolar en sitio atrófico único en maxila anterior usando hidroxiapatita y rhBMP-2

SUMMARY: Bone morphogenetic protein (rhBMP-2) is a powerful osteo-inductive growth factor widely used in bone reconstruction and both the vehicle used to administer it and the scaffold substrate could determine its success in clinical situations. The aim was to analyse the clinical behaviour of dental implants placed in single alveolar ridges with a horizontal deficiency in the maxillary anterior region that were reconstructed horizontally with rhBMP-2 and porous hydroxyapatite (HA). Inclusion criteria were both males and females, between the ages of 18 and 29 with single tooth loss of one upper incisor. Cone Beam Computed Tomography (CBCT) was used to take measurements prior to bone augmentation and again prior to the implant insertion. Surgery was carried out under local anaesthetic. In the primary procedure, bone substitute was introduced using porous HA and rhBMP-2; after 4 to 5 months, dental implant surgery was carried out and the implant placed; after 3 months of consolidation the provisional prosthesis was placed and then a definitive restoration was placed. Variables were analysed using the t-test with a p-value of < 0.05 in order to assess statistical significance. Thirteen subjects were included (6 females and 7 males). Bone augmentation resulted in a bone gain of 4.15mm (p=0.023), which was shown to be statistically significant. All of the grafts placed were successful and 13 implants were placed, using torques between 30 and 70N, without complications. For the final prostheses, 11 were screw retained and 2 were cemented in place. The horizontal bone augmentation using HA and rhBMP-2 is an efficient technique for single bone defects in the anterior maxillary area; clinical trials on a larger scale are needed to confirm these results.

RESUMEN: La proteína ósea morfogenética (BMP-2) es un potente osteoinductor utilizado ampliamente en técnicas reconstructivas; el vehículo de instalación es determinante en su evolución. El objetivo fue analizar el comportamiento clínico de implantes dentales instalados en rebordes alveolares únicos con deficiencia horizontal del sector anterior reconstruida horizontalmente con BMP-2 e hidroxiapatita (HA) porosa. Fueron incluidos sujetos de ambos sexos de entre 18 y 29 años, con pérdida dentaria unitaria a nivel de incisivos superiores. Se utilizó tomografía computadorizada para realizar mediciones en las etapas previa a la instalación del injerto y previo a la instalación del implante. Las cirugías fueron realizadas bajo anestesia local. En la primera intervención se realizó la instalación del injerto óseo utilizando HA porosa y BMP-2; después de 4 a 5 meses se realizó la instalación del implante dental; 3 meses después se realizó la conexión protésica y rehabilitación final. Las variables fueron estudiadas con la prueba t test considerando el valor de p< 0,05 para considerar significancia estadística. Trece sujetos fueron incluidos (6 mujeres y 7 hombres); con la reconstrucción ósea se obtuvo una ganancia ósea de 4,15mm (p=0.023) que fue estadísticamente significativo. No existió pérdida en ningún injerto realizado; se instalaron 13 implantes con torques entre 30 y 70N sin complicaciones; se realizaron prótesis fijas atornilladas en 11 casos y cementadas en 2 casos. La técnica con HA y BMP- 2 es eficiente para reconstruir defectos horizontales en perdidas unitarias del sector anterior maxilar; ensayos clínicos de mayor escala son necesarios para confirmar estos resultados.

Osteogenesis in vitro and bone defect repair in vivo of bone morphogenetic protein 9 composite collagen based bone repair material / 中国组织工程研究

BACKGROUND: Collagen-based bone repair materials have been widely used in clinical bone repair due to their excellent osteoconductivity, osteoinduction, degradation, mechanical properties and biocompatibility, but their osteoinduction effect is not as good as that of autogenous bone. OBJECTIVE: Bone morphogenetic protein 9 (BMP-9) was incorporated into collagen matrix to investigate its osteoinductive ability. METHODS: The plasmid DNA encoding BMP-9 was loaded into collagen-based bone repair materials to construct a composite of collagen-based bone repair materials. (1) In vitro experiment: The Sprague-Dawley rat adipose mesenchymal stem cells were inoculated with BMP-9 composite collagen-based bone repair material and collagen-based bone repair material. Cell proliferation was detected by MTT assay, and the cell adhesion was observed by scanning electron microscope. Cell adhesion was counted after DAPI staining. Alkaline phosphatase activity was detected by alkaline phosphatase kit. Osteopontin and osteocalcin mRNA expression levels were detected by qRT-PCR. (2) In vivo experiment: Bone defects of 3 mm in diameter and 3 mm in depth were made in the medial epicondyle of the right hind limb of 15 Sprague-Dawley rats. No material was implanted in the blank group. Collagen-based bone repair material was implanted in the control group. BMP-9 composite collagen-based bone repair material was implanted in the observation group. Samples were obtained at 8 weeks after surgery. Micro-CT examination and histological observation were performed. This study was approved by the Ethics Committee of Laboratory Animal Science Center of Nanchang University. RESULTS AND CONCLUSION: (1) In vitro experiment showed that there was no significant difference between the proliferation of adipose tissue mesenchymal stem cell cultured for 1, 3, 5, and 7 days in collagen-based bone repair materials compounded with BMP-9 (P > 0.05). (2) After co-culture in vitro for 24 hours, more cells and more pseudopodia appeared on the surface of collagen-based bone repair material compounded with BMP-9. The number and spreading of adherent cells were better than that of collagen-based bone repair material. (3) After 3 and 7 days of osteogenic induction in vitro, the cell alkaline phosphatase activity of the collagen-based bone repair material compounded with BMP-9 was higher than that of the collagen-based bone repair material (P < 0.05). After 7 days of osteogenic induction, osteopontin and osteocalcin mRNA expression was higher than that of collagen-based bone repair material (P < 0.05). (4) In vivo micro-CT examination showed that the amount of new bone formation in the observation group was higher than that in the control group and the blank group (P < 0.05), and that in the control group was higher than that in the blank group (P < 0.05). (5) In vivo hematoxylin-eosin staining and Masson staining showed that the bone repair effect of the observation group was better than that of the control and blank groups, and that in the control group was better than that in the blank group. (6) These findings confirmed that BMP-9 can further improve the bone induction properties of collagen-based bone repair materials.

Using allogeneic cortical graft preserved in glycerin as spacer in the advancement of tibial tuberosity in 34 dogs / Uso de enxerto cortical alogênico preservado em glicerina como espaçador no avanço da tuberosidade tibial em 34 cães

Cranial cruciate ligament is the main responsible for knee stability by preventing cranial tibial displacement regarding the femur. Deficiency in this ligament (CCLD) may cause subluxation of the tibia and dysfunction of the pelvic member due to overloading. Tibial osteotomies are among the more current surgical techniques for treating CCLD in dogs and they proportionate the dynamic stability by means of modifying bone geometry and the distribution of forces acting on the articulation. The objective of this work is to describe the use of the allogeneic cortical bone graft conserved in glycerin as a spacer on the tibial tuberosity advancement (TTA) for treating the CCLD. In order to do that, 34 dogs submitted to TTA surgery correction were evaluated, being 23 males (67.35%) and 11 females (32.35%). Surgical procedures happened from May 2011 to October 2015. Regarding the surgical procedure after osteotomy of the tibial tuberosity, a disk of allogeneic cortical disk, sawn wedge-hapsed, conserved in glycerin, proportions of 2x1mm was applied as spacer, enabling TTA. Advancements from 3 to 12 mm were executed, depending on the need of the patient. For animals with patella dislocation, trochleoplasty and TTA were executed in order to correct the deviation. The mean ± SD age of animals was 6.67±3.58 and weight was 15.16±12.97 kg. Mongrel dogs, Poodles and Yorkshire terriers were the most affected ones. From the 36 evaluated knees, 11 (30.56%) were associated with some traumatic process and in 25 (69.44%) there was no relation with previous trauma. From those wounds, 20 (55.56%) happened in the right limb and 16 (44.44%) in the left limb and two animals had CCLD bilaterally. Animals had continuous support, discreet drawer movement and negative tibial compression 15 days after surgery. At 30 days, 26 cases (72.22%) had firm support (FS); at 45 days, 24 cases (66 test at 7 and 67%) had FS and eight cases (22.22%) without claudication (WC). During subsequent radiographic evaluations the progressive incorporation of the graft and osteotomy union were observed. In this study, most of the diagnosed CCLD occurred in males diverging from results obtained by other authors that found greater frequency in females. Support without claudication it was observed in most of the cases of implants at 60 days. We concluded that the conserved allogeneic cortical bone graft was able to promote bone union in TTA of dogs with CCLD. None of the animals had signs of contamination, infection of the surgical wound or rejection related with the presence of the graft, demonstrated by the complete graft-bone incorporation observed early at 45 days in some animals. The glycerin was a good conservation medium for those fragments intended for grafting because, besides being of low cost, it kept bone fragments free of contamination, reducing antigenicity and preserving the functions of osteoinduction and osteoconduction. The possibility of molding the graft to the animal need is a characteristic favorable to executing the modified technique that could be molded according to the size of the animal, allowing perfect adaptation to the osteotomized local in different breeds. Intercurrences commonly observed in TTA with patellar dislocation, meniscal lesions, tibial crest fracture and displacement were not found in the animals of this study, probably due to the better distribution of forces between the pass screw in TT and the TTA plate confirming that it has good adaptation to the technique conferring to the modified TTA advantages regarding the conventional TTA.(AU)

O ligamento cruzado cranial é o principal responsável pela estabilidade do joelho, impedindo o deslocamento da tíbia cranial em relação ao fêmur. A deficiência neste ligamento (CCLD) pode causar subluxação da tíbia e disfunção do membro pélvico devido à sobrecarga. As osteotomias tibiais estão entre as técnicas cirúrgicas mais atuais para o tratamento de CCLD em cães e proporcionam a estabilidade dinâmica por meio da modificação da geometria óssea da distribuição das forças que atuam sobre a articulação. O objetivo desse estudo é descrever o uso do enxerto ósseo cortical alogênico conservado em glicerina como espaçador no avanço da tuberosidade tibial (TTA) para o tratamento do CCLD. Para isso, 34 cães submetidos à cirurgia de TTA foram avaliados, sendo 23 machos (67,35%) e 11 fêmeas (32,35%). Os procedimentos curúrgicos aconteceram entre maio de 2011 e outubro de 2015. Com relação ao procedimento cirúrgico após a osteotomia da tuberosidade tibial, um disco alogênico cortical, em forma de cunha serrada, conservado em glicerina com proporções de 2 x 1mm foi aplicado como espaçador possibilitando a TTA. Avanços de 3 a 12mm foram executados, dependendo da necessidade do paciente. Para animais com luxação da patela, realizou-se a trocleoplastia e a TTA para a correção do desvio. A idade média dos animais foi de 6,67±3,58 anos e pesos médios de 15,16±12,97kg. Cães sem raça definida, Poodles e Yorkshire Terriers foram os mais afetados. Dos 36 joelhos avaliados, 11 (30,56%) foram associados a algum processo traumático e em 25 (69,44%) não havia nenhuma relação com um trauma prévio. Dos ferimentos, 20 (55,56%) aconteceram no membro direito e 16 (44,44%) no esquerdo, sendo que dois animais apresentavam CCLD bilateralmente. Os animais tiveram suporte contínuo, discreto movimento de gaveta e compressão tibial negativa 15 dias após a cirurgia. Aos 30 dias, 26 casos tinham suporte firme (FS); aos 45 dias, 24 casos tinham FS e oito casos sem claudicação (WC). Durante avaliações radiográficas subsequentes, observou-se a incorporação progressiva da união do enxerto e da osteotomia. Neste estudo, a maior parte do CCLD diagnosticado ocorreu em machos, divergindo dos resultados obtidos por outros autores que encontraram maior frequência em fêmeas. Suporte sem claudicação foi observado na maioria dos casos de implantes aos 60 dias. Foi concluído que o enxerto ósseo cortical alogênico conservado foi capaz de promover a união óssea na TTA de cães com CCLD. Nenhum dos animais apresentou sinais de contaminação, infecção da ferida cirúrgica ou rejeição relacionada à presença do enxerto, demonstrada pela incorporação completa do enxerto ósseo observada precocemente aos 45 dias em alguns animais. A glicerina foi um bom meio de conservação para os fragmentos destinados à enxertia porque, além do menor custo, manteve os fragmentos ósseos livres de contaminação, reduzindo a antigenicidade e preservando as funções de osteoindução e osteocondução. A possibilidade de moldagem do enxerto à necessidade do animal é uma característica favorável à execução da técnica modificada que pode ser moldada de acordo com o tamanho do animal, possibilitando perfeita adaptação ao local osteotomizado em diferentes raças. Intercorrências comumente observadas na TTA com luxação patelar, lesões meniscais, fratura da crista tibial e deslocamento não foram encontradas nos animais deste estudo, provavelmente devido à melhor distribuição de forças entre a passagem do parafuso no TT e a placa do TTA, confirmando que tem boa adaptação à técnica conferindo às vantagens da TTA modificada em relação à TTA convencional.(AU)

Experimental study of demineralized dentin matrix on osteoinduction and related cells identification / 华西口腔医学杂志

OBJECTIVE@#The aim of this study was to explore the theoretical framework of cells and the forms of osteogenesis in the mechanism by which demineralized dentin matrix (DDM) induces osteogenesis.@*METHODS@#A total of 24 New Zealand rabbits were used in this study. A total of 4 erector spinae bags were created in each animal. A total of 3 erector spinae bags were implanted with DDM by random selection, whereas the remaining one erector spinae bag was not implanted with DDM. The rabbits were sacrificed after 1, 2, 3, 4, 8, 12, 16, and 20 weeks, and the samples were obtained. The samples were examined by hematoxylin-eosin (HE), tartrate-resistant acid phosphatase (TRAP), and immunohistochemical staining to identify the mesenchymal stem cells, osteoblasts, chondrocytes, and osteoclasts.@*RESULTS@#The results of HE staining showed that in the third week, cartilage- and bone-like matrices, as well as the osteoblast-like cells, were observed. The results of immunohistochemical staining showed that the expressions of CD44, alkaline phosphatase (ALP), and collagen Ⅱ were statistically significant 
(P<0.05).@*CONCLUSIONS@#DDM has good histocompatibility and osteoinduction. In addition, induced ectopic osteogenesis mode mainly occurs in the endochondral bone.

Bioinductive biomaterials for periodontal regeneration / 中华口腔医学杂志

Periodontal regeneration is a great challenge faced by clinical treatment, and traditional therapies are not effective in regenerating periodontal tissues. Bioinductive biomaterials are materials that are able to induce the regeneration of damaged tissues, and they can induce the migration of stem cells or precursor cells to the defect site to proliferate, differentiate for tissue regeneration. For example, the osteoinductive biomaterials are able to induce regeneration of alveolar bone. In this review, the writer combined the recent advances of both clinical and scientific field, and summarize the application of bioinductive biomaterials includingautogenous bone, human derived growth factors, enamel matrix proteins and platelet-rich fibrin.

Efecto osteoinductor del mineral trióxido agregado versus el cemento Portland tipo I sobre lesiones óseas mandibulares

Introducción: Estudio experimental donde se procuró determinar el efecto osteoinductor del mineral trióxido agregado (MTA) versus el cemento Portland tipo I sobre lesiones óseas mandibulares. Metodología: Se emplearon 12 conejos machos de la raza New Zealand de 3 meses de edad, los cuales fueron divididos en 4 grupos iguales. Todos los conejos fueron anestesiados utilizando pentobarbital sódico. Se procedió a la incisión en la piel mandibular para exponer el hueso sobre el que se realizó 3 cavidades de 3mm cada una. En una cavidad se colocó MTA, en otra cemento Portland y en la tercena ninguna pasta. Se procedió al sacrificio de los grupos experimentales a la 1era, 2da, 3era y 4ta semana respectiva y se evaluó las muestras obtenidas de las áreas quirúrgicas mediante conteo de osteocitos y osteoblastos. Resultados: Tanto el MTA como el cemento Portland poseen la misma capacidad osteoinductiva en la 1era, 2da y 3era semana (p>0,05). Sin embargo, en la 4ta semana el MTA tuvo mayor capacidad osteoinductora al estimular mayor número de osteoblastos que el cemento Portland (p=0,024). Conclusiones: El MTA y el cemento Portland tipo I mostraron similar efecto osteoinductor durante las 3 primeras semanas de evaluación. El MTA demostró mayor efecto osteoinductor durante la cuarta semana de valoración.

Introduction: An experimental study was carried out to determine the osteoinductive effect of Mineral Trioxide Aggregate (MTA) versus Portland Cement type I on mandibular bone lesions. Methodology: Twelve 3-month-old male New Zealand rabbits were divided into 4 equal groups. All rabbits were anesthetized using Pentobarbital. An incision of the mandibular skin was performed to expose the bone on which 3 cavities of 2mm each one were made. In one cavity MTA was placed, in another Portland Cement type I and the third remained empty. The experimental groups were sacrificed at the 1st, 2nd, 3rd and 4th respective weeks and evaluated histologically by counting osteocytes and osteoblasts. Results: Both MTA and Portland cement have the same osteoinductive capacity in the 1st, 2nd and 3rd week (0.05

The osteointergration and osteoinduction of titanium implant with nHA/BG gradient coating in rabbits / 实用口腔医学杂志

Objective: To investigate the osteointegration and osteoinduction of nano hydroxyapatite/bioglass ( nHA/BG ) gradient nanofilm on the surface of titanium ( Ti) prepared by hypotherm sintering and plastic deformation. Methods:Hypotherm sintering was used to produce nHA/BG gradient coating followed by soaking in the simulated body fluid. Ti implants with gradient coatings were planted in femoral condyles at one side of 12 New Zealand rabbits and the untreated Ti implants were planted at the other side as the controls. 1, 3 and 6 months after implantation, the animals were sacrificed after X-ray examination and the tissues around the implants from the 3 month group were used for the preparation of hard tissue section and ground section. New bone formation was observed by tetracycline fluorescence staining. Von Gieson staining was used to observe the osteointegration at the interface between bone and im-plant. Results:The gradient coatings were porous and composed of irregular rod-like nano-HA crystals. Animal study showed well es-tablished osteointegration between the gradient coating and more novel bone was found around the implants with gradient coatings. Conclusion:Osteointegration and ostioinduction of Ti implant can be enhanced by nanostructured surface with gradient coatings of nHA/BG.

The experimental study on osteogenic potential of goat adipose derived stromal cells induced by PRP with rhBMP-2 / 中华显微外科杂志

Objective To evaluate the feasibilty of modified osteogenic culturing of goat adipose derived stromal cells(ADSCs).Methods From March,2013 to September,2014,platelet-rich plasma(PRP) was made from goat autogenous vein blood,and abodimoneal fat was aspirated,following aspetic procedure,primary and series passage of ADSCs was established.Osteoinduced ADSCs was carried out according to following group:combinative osteoinduction group(PRP+rhBMP-2 +ADSCs),growth factor group(rhBMP-2+ADSCs),conventional inductive group(dexamathasone+ascorbic acid + ADSCs) and noninductive group(blank group).Converted microscope was used to observe cellular pattern,cell activity,osteocalcin and collagen type Ⅰ level were detected at 1,3,5,7,9,13,17,21 days.Red Alazarin and Von Kossa staining were also assayed at different interval.Results Under observation of converted microscopy,remarkable cell proliferation with abundant ECM was noticed in combinative osteoinduction group,compared with other groups,level of cell activity,osteocalcin,collagen type Ⅰ [(0.82 ± 2.19)AU,(79.82 ± 1.36)U/L at,(78.51 ± 4.32)μ g/ml at]were significantly higher than other groups (P ＜0.05),and remarkable ALP expression and calcifed nodules were also seen.Conclusion PRP can enhance the inductive effect of rhBMP-2,and combinative osteoinduction procedure acts as a satisfactory culturing method.

Cranial bone regeneration according to different particle sizes and densities of demineralized dentin matrix in the rabbit model / 대한악안면성형재건외과학회지

BACKGROUND: The objective of this study was to place bone graft materials in cranial defects in a rabbit model and compare their bone regenerating ability according to the size and density of demineralized dentin matrix (DDM). METHODS: We selected nine healthy male rabbits that were raised under the same conditions and that weighed about 3 kg. Two circular defects 8 mm in diameter were created in each side of the cranium. The defects were grafted with DDM using four different particle sizes and densities: 0.1 mL of 0.25- to 1.0-mm particles (group 1); 0.2 mL of 0.25- to 1.0-mm particles (group 2); 0.1 mL of 1.0- to 2.0-mm particles (group 3); and 0.2 mL of 1.0- to 2.0-mm particles (group 4). After 2, 4, and 8 weeks, the rabbits were sacrificed, and bone samples were evaluated by means of histologic, histomorphometric, and quantitative RT-PCR analysis. RESULTS: In group 1, osteoblast activity and bone formation were greater than in the other three groups on histological examination. In groups 2, 3, and 4, dense connective tissue was seen around original bone even after 8 weeks. Histomorphometric analysis of representative sections in group 1 showed a higher rate of new bone formation, but the difference from the other groups was not statistically significant. RT-PCR analysis indicated a correlation between bone formation and protein (osteonectin and osteopontin) expression. CONCLUSIONS: DDM with a space between particles of 200 μm was effective in bone formation, suggesting that materials with a small particle size could reasonably be used for bone grafting.

Histologic study of bone-forming capacity on polydeoxyribonucleotide combined with demineralized dentin matrix / 대한악안면성형재건외과학회지

BACKGROUND: This study examined the osteoinductive activity of demineralized dentin matrix (DDM) from human and polydeoxyribonucleotide (PDRN) for nude mice. METHODS: Twenty healthy nude mice, weighing about 15~20 g, were used for the study. DDM from human and PDRN were prepared and implanted subcutaneously into the dorsal portion of the nude mice. The nude mice were sacrificed at 1, 2, and 4 weeks after grafting and evaluated histologically by hematoxylin-eosin and Masson's trichrome staining. The specimens were also evaluated via a histomorphometric study. RESULTS: The DDM and PDRN induced new bone, osteoblasts, and fibroblasts in soft tissues. The histological findings showed bone-forming cells like osteoblasts and fibroblasts at 1, 2, and 4 weeks. New bone formation was observed in the histomorphometric study. In particular, the ratio of new bone formation was the highest at 2 weeks compared with the first week and fourth week. CONCLUSIONS: In this study, we showed that the PDRN used in this experimental model was able to induce bone regeneration when combined to the DDM.

Morphological study of bone cranial in athymic mice / Estudio morfológico de hueso craneal en ratones atímicos

The aim of our research was to create an osteogenic unit in the skulls of athymic mice; however, the first challenge we faced was to find sufficient and adequate data that would allow us to determine the morphological, immunohistochemical and microtopographical characteristics that could be used as normality standards in athymic mice skulls and, hence, a reference in the event of achieving the formation of de novo bone using the osteogenic unit we proposed. Knowing the normal bone morphology in the skull of athymic mice was a necessary precondition to develop subsequently an osteogenic unit possessing the Osteogenesis, Osteoinduction and Osteoconductivity that could be compared versus those in the normal bone during its formations and remodeling processes. Therefore, we conducted a pilot study to determine bone morphological characteristics in the skull of athymic mice by means of specific histological staining: hematoxylin-eosin and Von Kossa, for osteoid tissue and mineralized bone, and Masson Tri-chrome for ossified areas. We also use immunohistochemistry to detect bone formation markers: alkaline phosphatase resulting from osteoblastic activity stimulation, type 1 collagen a bonematrix structural protein; Osteopontine, a protein specifically synthesized by osteoblasts that favors cell proliferation and remodeling in bone defects; Osteocalcine, a peptide hormone produced by osteoblasts during bone formation; and, Runx 2, a transcription factor expressed by stem cells which stimulates bone differentiation. Likewise, we used electron microscopy on the newly formed tissue to determine the presence of organic deposits, such as calcium, phosphate and magnesium in bone tissue.

Propusimos la realización de una unidad osteogénica a desarrollar en cráneo de ratones atímicos, Sin embargo, nos enfrentamos al reto de encontrar datos que nos determinaran cuales eran las características morfológicas, inmunohistoquímicas y micro-topográficas del cráneo de estos ratones atímicos, que nos sirvieran como referencia de normalidad y tener un punto de comparación, en caso de que pudiéramos lograr la formación de hueso de novo, a partir de la unidad osteogénica que propusimos. El objetivo, de conocer la morfología del hueso normal de cráneo de ratones atímicos, fue desarrollar posteriormente una unidad osteogénica que reuniera las características de Osteogénesis, Osteoinducción y Osteoconducción, y, compararlas contra las que tiene dicho hueso normal durante su proceso de formación y remodelación. Así, realizamos un estudio piloto donde establecimos características morfológicas de hueso del cráneo de ratones atímicos, a través de tinciones histológicas específicas, con hematoxilina-eosina y von Kossa para buscar tejido osteoide y hueso mineralizado y Tricrómico de Massón para observar zonas osificadas. Además, analizamos el tejido óseo a través de inmunohistoquímica, con la finalidad de buscar marcadores de formación ósea como fosfatasa alcalina que es resultado del estímulo de la actividad osteoblástica; colágena 1, la cual es una proteína estructural de la matriz ósea; osteopontina, proteína sintetizada específicamente por osteoblastos que favorece la proliferación celular y la remodelación en defectos óseos; osteocalcina hormona peptídica producida por los osteoblastos durante la formación ósea y Runx 2 Factor de transcripción expresado por las células progenitoras que estimula la diferenciación ósea. Además, sometimos el tejido óseo a microscopía electrónica para determinar la presencia de depósitos de compuestos como calcio, fósforo y magnesio.

Preparation and ectopic osteogenesis in vivo of scaffold based on new synthetic biodegradable hydrogel loaded with synthetic BMP-2-derived peptide / 国际生物医学工程杂志

Objective To prepare P24/PTMC11-F127-PTMC11 hydrogel,to study the in vitro release profile and to observe ectopic bone formation in p24 peptide incorporated PTMC11-F127-PTMC11 hydrogel.Methods Corresponding weight powder of p24 peptide was infunded into tubes of PTMC11-F127-PTMC11 solution with concentrations of 16％,20％ and 25％.Release profiles of P24 peptide in different concentration PTMC11-F127-PTMC11 hydrogel were measured in vitro by BCA assay.P24/PTMC11-F127-PTMC11 hydrogel was implanted into each rat's erector muscle of spine,and the implanted gel was detected by hematoxylin and eosin stain (HE).Results PTMC11-F127-PTMC11 hydrogel showed sustained slow release for the whole process after the initial burst release.With the increase of concentration in PTMC11-F127-PTMC.hydrogel,the initial burst release was reduced significantly.Ectopic bone formation was observed by computed tomography in p24 peptide incorporated PTMC11-F127-PTMC11 hydrogel after four weeks.Bone trabeculae surround the P24/PTMC11-F127-PTMC11 hydrogel was observed at forth week by hematoxylin and eosin stain.The bone trabeculae became thicker from sixth week.Conclusion Delayed release of peptide from the hydrogel was mainly controlled by disintegration of hydrogel and a satisfactory release profile was observed.These results suggest that the p24-loaded PTMC11-F127-PTMC11 hydrogel remmns active of p24 at the implanted site,continuously induce differentiation of osteoblast precursor cells into osteoblasts,and activate osteoblasts to promote ectopic calcification.

Avaliação microscópica da propriedade biológica e da biocompatibilidade do osso composto e do osso integral de origem bovina / Microscopic evaluation of biological properties and biocompatibility of bovine, non-demineralized lyophilized and composite bovine bones

Este trabalho teve como objetivo avaliar a propriedade biológica e a biocompatibilidade do osso composto e do osso integral de origem bovina, implantados em cavidades ósseas de tíbias e no plano subcutâneo de ratos. Foram utilizados 24 ratos sacrificados após 15 e 45 dias. Os resultados do exame microscópico das peças obtidas do subcutâneo mostraram aos 15 dias uma reação inflamatória ao redor das partículas do material com presença de células gigantes e, aos 45 dias, observou-se diminuição da reação inflamatória e presença de tecido conjuntivo fibroso ao redor das partículas com a presença de células gigantes. Não houve indícios de formação óssea ectópica nas peças histológicas obtidas da tíbia; foi possível observar maior atividade de neoformação óssea no grupo controle aos 15 dias com 42,8% da cortical externa sendo neoformada em comparação aos 22,6% do grupo Orthogen e 25% do grupo GenMix. Aos 45 dias havia 62,5% de neoformação óssea no grupo controle, 26% no grupo Orthogen e 35% no grupo GenMix. Pôde-se concluir que o osso composto e o osso integral de origem bovina são materiais biocompatíveis, possibilitam a neoformação óssea por suas qualidades osseocondutivas e não induzem à formação de osso ectópico...

The aim of this study was to evaluate the biological properties and biocompatibility of bovine non-demineralized lyophilized and composite bones implanted in tibiae bone cavities and at the subcutaneous level. Twenty-four rats were used and sacrificed 15 and 45 days later. At the subcutaneous level, after 15 days an inflammatory reaction was seen around biomaterial particles with the presence of giant cells and at 45 days fibrous connective tissue had also developed. No signs of ectopic bone formation were observed at tibiae regions; more bone neoformation was observed at the control group (15 days) with 42.8% of the outer cortex layer against 22.6% at Orthogen and 25% at GenMix groups. At 45 days, correspondent values for bone neoformation were 62.5% at control, 26% at Orthogen, and 35% at GenMix groups, respectively. It can be concluded that both materials tested were biocompatible aiming to bone neoformation by their osteoconductive properties with no ectopic formation sites observed...

Three-year Follow-up after Autogenous and Xenogenic Jaw Bone Grafts

Clinical application of auto-tooth bone graft material

INTRODUCTION: Auto-tooth bone graft material consists of 55% inorganic hydroxyapatite (HA) and 45% organic substances. Inorganic HA possesses properties of bone in terms of the combining and dissociating of calcium and phosphate. The organic substances include bone morphogenetic protein and proteins which have osteoinduction capacity, as well as the type I collagen identical to that found in alveolar bone. Auto-tooth bone graft material is useful as it supports excellent bone regeneration capacity and minimizes the possibility of foreign body reaction,genetic diseases and disease transmission. MATERIALS AND METHODS: Implant placement combined with osteoinductive regeneration,preservation of extraction socket, maxillary sinus augmentation, and ridge augmentation using block type,powder type, and block+powder type autobone graft materialwere performed for 250 patients with alveolar bone defect and who visited the Department of Oral and Maxillofacial Surgery, College of Dentistry, Dankook University from September 2009 to August 2011. RESULTS: Clinical assessment: Among the 250 patients of auto-tooth bone graft, clinical assessment was performed for 133 cases of implant placement. The average initial stabilization of placed implants was 74 implant stability quotient (ISQ). Radiological assessment: The average loss of crestal bone in the mandible as measured 6 months on the average after the application of prosthesis load was 0.29 mm, ranging from 0 mm to 3.0 mm. Histological assessment: In the histological assessment, formation of new bone, densified lamellated bone, trabecular bones, osteoblast, and planting fixtures were investigated. CONCLUSION: Based on these results, we concluded that auto-tooth bone graft material should be researched further as a good bone graft material with osteoconduction and osteoinduction capacities to replace autogenous bone, which has many limitations.

Guided Bone Regeneration Using a Putty-type Demineralized Bone Matrix: Case Report

Bone substitute in the repair of the post-extraction alveolus

In recent years there has been increasing interest in the choice of the best material for bone substitutes. Experimental models enable estimation of biological potential, efficacy and safety of a biomaterial before its clinical application. The aim of this study was to evaluate the response of a bone substitute, UNC bone matrix powder (MOeP-UNC), for repairing the postextraction alveolus in Wistar rats. Rats' first lower molars were extracted. The right alveoli were filled with MOeP-UNC hydrated with physiological saline (Experimental Group, EG), and the left alveoli were used as Control Group (CG). Thirty days after extraction, the animals were killed and the samples processed. Histological sections were made in vestibular- lingual direction at the level of the mesial alveolus of the first inferior molar (Guglielmotti et al. J. Oral Maxillofac. Surg. 1985;43(5):359-364). Repair of the alveoli at 30 days after extraction was evaluated histologically. Repair of the alveolus was optimum in the control group at 30 days, and the EG showed presence of MOeP-UNC particles in close contact with newly formed bone tissue (osseointegration). In the experimental model used, at 30 days post-surgery, the MOeP-UNC particles integrate compatibly with newly formed bone tissue.

En los ultimos anos se ha incrementado el interes por la eleccion del material mas adecuado como sustituto oseo. Los modelos experimentales permiten estimar el potencial biologico, la eficacia y seguridad de un biomaterial, previo a su aplicacion clinica. El objetivo del presente estudio fue evaluar la respuesta de un sustituto oseo, matriz osea-UNC en polvo (MOeP-UNC), en la reparacion alveolar post-exodoncia en ratas Wistar. Se realizo la exodoncia de los primeros molares inferiores. En los alveolos derechos se coloco MOeP-UNC hidratada con solucion fisiologica (Grupo Experimental, GE). Los alveolos izquierdos, fueron utilizados como Grupo Control (GC). A los 30 dias post-exodoncia los animales fueron sacrificados y las muestras obtenidas se procesaron, se realizaron cortes histologicos en sentido vestibulo-lingual a la altura del alveolo mesial del primer molar inferior (Guglielmotti et al. J Oral Maxillofac Surg. 1985;43(5):359-364). Se realizo la evaluacion histologica de la reparacion de los alveolos a los 30 dias post cirugia. El grupo control presento una optima reparacion alveolar a los 30 dias y en el GE se evidencio la presencia de las particulas de MOeP-UNC en intimo contacto con el tejido oseo neoformado (oseointegracion). En el modelo experimental utilizado, a los 30 dias post-cirugia las particulas MOePUNC se integran de manera compatible con el tejido oseo neoformado.

Morphometric analysis on bone formation effect of beta-TCP and rhBMP-2 in rabbit mandible

INTRODUCTION: This study was to assess the effectiveness of new bone formation and regeneration by using a rhBMP-2 and beta-TCP as a carrier in rabbits' mandible. MATERIALS AND METHODS: The mandibles of 36 rabbits were exposed and cortical bone was penetrated for this study. The experimental subjects were divided into 3 groups each 12 rabbits; control group, experimental group 1, and experimental group 2. Control group had the defect itself without any treatment, in the experimental group 1, beta-TCP only was grafted, and in the experimental group 2, rhBMP-2 soaked in beta-TCP was grafted. The rabbits were sacrificed after 1, 2, 3, 4, 6, and 8weeks, and new bone formation area was examined and measured for bone quantitative and qualitative analysis with light, fluorescent and polarized microscopy. RESULTS: In the experimental group 1, new bone formation from the adjacent host bone was made by osteoconduction, and in the experimental group 2, direct new bone formation by osteoinduction of rhBMP-2 as well as new bone formation by osteoconduction of beta-TCP were observed. CONCLUSION: rhBMP-2 of experimental group 2 is very effective in the bone formation in early 2weeks and bone remodelling from 3weeks.

Osteoinductive Potential of the rhBMP-2 in Soft Tissues / Potencial Osteoinductor de la rhBMP-2 en Tejidos Blandos

The rhBMP-2 is an osteoinductive protein used in the reconstructive with the objective to create newly formed bone. The aim of this study was to confirm the rhBMP-2 osteoinductive capability, when implanted in soft tissues. The results showed that the protein used in this study is highly osteoinductive.

La proteína oseoinductora rhBMP-2 es usada en cirugías reconstructoras, con la finalidad de crear tejido óseo neoformado. El objetivo de este estudio fue confirmar la capacidad oseoinductora de la rhBMP-2, cuando fue implantada en tejidos blandos. Los resultados mostraron que la proteína usada en este estudio es altamente oseoinductora.