Effects of bleaching using 10% carbamide peroxide with calcium or amorphous calcium phosphate on enamel mineral content and hardness / Asociación entre la severidad de Hipomineralización Incisivo-Molar (HIM) y lesiones cavitadas de caries en escolares

ABSTRACT This study evaluated enamel mineral content and surface microhardness before and after bleaching treatment using 10% carbamide peroxide (CP) containing calcium (Ca) or amorphous calcium phosphate (ACP). Thirty-six bovine slabs were randomly allocated into 3 groups (n = 12) according to bleaching treatment: G1 - Opalescence PF 10% (CP), G2 -NiteWhite ACP (CP+ACP), and G3 - Opalescence PF (10%) with calcium (CP+CA). The bleaching agent was applied on enamel surface for 6 h/day over a period of 21 days. Enamel surface was evaluated by Knoop microhardness (KNH) and micro energy-dispersive X-ray fluorescence spectrometry (p-EDXRF) at baseline and at after bleaching treatment. Data were statistically analyzed by repeated measures ANOVA and Tukey's test (a = 0.05). There was a significant decrease in microhardness after bleaching treatments for all study groups, but no difference between bleaching gels. There was no difference in the Ca/P ratio measured by p-EDXRF for all groups at the study times, but the mean value was lower in group CP+CA than in group CP+ACP. Group CP was similar to both CP+ACP and CP+CA. It can be concluded that enamel microhardness decreased after the bleaching process, regardless of the presence of calcium or ACP, but there was no significant change in the Ca/P ratio of enamel after bleaching for each tested gel. This indicates that the bleaching gels have erosive potential, causing softening of enamel without promoting surface loss, regardless of the presence of calcium of ACP ions.

RESUMO Este estudo avaliou o conteúdo mineral do esmalte e a microdureza superficial antes e após o tratamento clareador, utilizando peróxido de carbamida 10% (PC) contendo cálcio (Ca) ou fosfato de cálcio amorfo (ACP) em sua composigao. Trinta e seis espécimes de esmalte bovino foram alocados aleatoriamente em 3 grupos (n = 12) de acordo com os tratamentos clareadores: G1 - Opalescence PF 10% (CP), G2 -NiteWhite (CP+ACP); e G3 - Opalescence PF (10%) com cálcio (CP + CA). O agente clareador foi aplicado na superficie do esmalte por 6 h/dia por um periodo de 21 dias. A superficie do esmalte foi avaliada por microdureza Knoop (KNH) e espectrometria de fluorescencia de raios X micro-dispersiva (p-EDXRF) no inicio e após o tratamento clareador. Os dados foram analisados estatisticamente pelo teste ANOVA de medidas repetidas e Tukey (a = 0,05). Houve uma diminuigao significativa da microdureza após os tratamentos clareadores para todos os grupos estudados, mas nao houve diferenga entre os diferentes géis. Nao houve diferenga da relagao Ca/P mensurada por p-EDXRF para todos os grupos nos tempos estudados; no entanto, o grupo CP+CA apresentou menor valor comparado ao grupo CP+ACP. O grupo CP foi similar aos grupos CP+ACP e CP+CA. Portanto, pode-se concluir que houve redugao significativa da microdureza do esmalte após o clareamento, independente da presenga de cálcio ou APC na composigao dos géis, embora nao tenha havido alteragao significando na relagao Ca/P do esmalte após o clareamento. Isto indica um potencial erosivo dos géis clareadores, causando o amolecimento sem perda da estrutura do esmalte, independente da presenga dos íons cálcio e ACP.

A comparative study of the biocompatibility of two root-end filling materials in rat connective tissue / Estudio comparativo de la biocompatibilidad de dos materiales de retro-obturación en tejido conectivo de rata

The aim of the present study was to examine the short-term biocompatibility of Endosequence Root Repair Material (ERRM) paste and white Mineral Trioxide Aggregate MTA by implanting them into polyethylene tubes in the subcutaneous connective tissue of rats. twenty five male Wistar rats, 3-4 months old, weighing 300-350 g, were used. The tubes were implanted dorsally into the subcutaneous connective tissues of the rats. Five animals were sacrificed at five examination time points: 1, 3, 5, 7 and 15 days. The connective tissues containing the implants were excised. These sections were studied qualitatively and quantitatively using a light microscope. An average value for each group was obtained by averaging the sum of all inflammatory cells counted in 10 randomly selected, separate areas. For the ERRM group: There was a significant increase in the number of inflammatory cells on days 1-3 and on days 5-7 (P ≤ 0.003 and P ≤ 0.024). In the WHITE MTA group, the mean values of the sum of the inflammatory cells during the periods 1-3 days and 5-7 days were statistically significant (P ≤ 0.001 and P ≤ 0.044, respectively) and the XILOPERCHA group: Difference was observed significant in the value of the sum of inflammatory cells during the period of 3-5 days (P ≤ 0.05). According to the results it can be concluded that both, ERRM as MTA, caused an inflammatory reaction, which decreased over time; suggesting that both materials are biocompatible; showing however the presence of a higher organization of collagen fibers around the implants of ERRM.

El objetivo del presente estudio fue evaluar la biocompatibilidad a corto plazo de Material de Reparación de la Raíz Endodóntica (MRRE) y el agregado de trióxido mineral (AgTM), implantándolos dentro de tubos de polietileno en el tejido conectivo subcutáneo de ratas. Se usaron 25 ratas Wistar macho, de 3-4 meses de edad, con peso de 300 a 350 g. Los tubos fueron implantados en el tejido conectivo subcutáneo del dorso de las ratas. Cinco animales fueron sacrificados en cada uno de los siguientes períodos de tiempo: 1, 3, 5, 7, y 15 días. El tejido conectivo con los implantes fue escindido y seccionado. Los cortes se evaluaron cualitativa y cuantitativamente mediante microscopio óptico. Se obtuvo un valor para cada grupo resultado al promediar la suma de las células inflamatorias contadas en 10 áreas separadas seleccionadas aleatoriamente. Para el grupo de MRRE; hubo un incremento significativo en la cantidad de células inflamatorias entre los días 1-3 y 5-7 (p ≤ 0,003 y p ≤ 0,024). En el grupo de AgTM blanco, los valores promedio de la suma de células inflamatorias entre los períodos 1-3 días, y 5-7 días mostraron ser estadísticamente significativos (p≤ 0,001 y p ≤ 0,044 respectivamente) y en el grupo control de Xilopercha se observó diferencia significativa entre los valores de la suma de células inflamatorias entre los períodos de 3-5 días (P ≤ 0,05). De acuerdo a los resultados, puede concluirse que ambos materiales, AgTM y MRRE causaron una reacción inflamatoria que disminuyó a través del tiempo, sugiriendo que ambos materiales son biocompatibles; mostrando sin embargo una mayor organización de fibras colágenas alrededor de los implantes de MRRE.

Demineralized bone matrix and calcium-phosphate cement in bone regeneration in rats

Abstract Purpose: To compare bone regeneration in critical-sized defects in rat calvarium using demineralized bone matrix and calcium phosphate cement. Methods: Thirty Wistar rats were divided into 3 groups of 10 animals each. Two defects of 5-mm were made in the parietal bones of each animal. Group I had calcium phosphate cement placed in the experimental defect, Group II had filled with demineralized bone matrix and Group III had with the combination of the matrix and cement in equal parts. All animals had one defect left unfilled to serve as controls. Five animals in each group were sacrificed at 4 and 8 weeks. Histomorphometric analysis was used to quantify the amount of new bone within the defects. Results: The results showed that demineralized bone matrix-treated defects had significantly more new bone at 4 weeks compared to calcium phosphate cement-treated defects (p=0.03) and also had significantly more new bone at 8 weeks compared to unfilled defects (p=0.04). Conclusions: The demineralized bone matrix was superior to calcium phosphate cement in bone regeneration. It seems that calcium phosphate cement acted by inhibiting the osteogenesis when associated with a demineralized bone matrix and this combination should not be recommended.

Repair of bone defects using adipose-derived stem cells combined with alpha-tricalcium phosphate and gelatin sponge scaffolds in a rat model

Abstract Objectives This study aimed to evaluate the potential of adipose-derived stem cells (ASCs) combined with a modified α-tricalcium phosphate (α-TCP) or gelatin sponge (GS) scaffolds for bone healing in a rat model. Material and Methods Bone defects were surgically created in the femur of adult SHR rats and filled with the scaffolds, empty or combined with ASCs. The results were analyzed by histology and histomorphometry on days seven, 14, 30, and 60. Results Significantly increased bone repair was observed on days seven and 60 in animals treated with α-TCP/ASCs, and on day 14 in the group treated with GS/ASCs, when compared with the groups treated with the biomaterials alone. Intense fibroplasia was observed in the group treated with GS alone, on days 14 and 30. Conclusions Our results showed that the use of ASCs combined with α-TCP or GS scaffolds resulted in increased bone repair. The higher efficacy of the α-TCP scaffold suggests osteoconductive property that results in a biological support to the cells, whereas the GS scaffold functions just as a carrier. These results confirm the potential of ASCs in accelerating bone repair in in vivo experimental rat models. These results suggest a new alternative for treating bone defects.

Repair of Surgical Bone Defects Grafted with Hydroxylapatite + ß-TCP and Irradiated with λ=850 nm LED Light

The treatment of bone loss due to different etiologic factors is difficult and many techniques aim to improve the repair, including a wide range of biomaterials and recently, photobioengineering. This work aimed to assess by histological analysis the repair of bone defects grafted with biphasic synthetic micro-granular HA + β-TCP associated with LED phototherapy. Forty rats were divided into 4 groups (Clot, LED, Biomaterial and LED + Biomaterial) each subdivided into 2 subgroups according to the time of animal death (15 and 30 days). Surgical bone defects were prepared on the femur of each animal with a trephine drill. In animals of the Clot group the defect was filled only by blood clot, in the LED group the defect filled with the clot was further irradiated. In the animals of Biomaterial and LED + Biomaterial groups the defect was filled by biomaterial and the last one was further irradiated (λ=850±10 nm, 150 mW, Φ ~ 0.5 cm2, 20 J/cm2 - session, 140 J/cm2- treatment) at 48-h intervals for 2 weeks. Following animal death, samples were taken and analyzed by light microscopy. Using the degree of maturation of the bone by assessment of the deposition/organization of the basophilic lines in the newly formed bone tissue, the LED + Biomaterial group was the one in a more advanced stage of bone repair process at the end of the experiment. It may be concluded that the use of LED phototherapy was effective in positively modulating the process of bone repair of bone defects in the femur of rats submitted or not to biomaterial grafting.

O tratamento de perdas ósseas devido a diferentes fatores etiológicos é difícil e muitas técnicas têm por objetivo melhorar o reparo incluindo o uso de uma ampla gama de biomateriais e, recentemente, a fotobioengenharia. Este trabalho teve como objetivo avaliar, por meio de análise histológica, o reparo de defeitos ósseos enxertados com HA bifásica micro-granular sintética + β -TCP associada à fototerapia LED. Quarenta ratos foram divididos em quatro grupos (Clot, LED, Biomaterial e LED + Biomaterial), subdivididos no dois subgrupos de acordo com o momento da morte (15 e 30 dias). Defeitos ósseos cirúrgicos foram criados em um fêmur de cada animal com uma broca trefina. Em animais do grupo coágulo, o defeito foi preenchido apenas pelo coágulo sanguíneo, no grupo de LED o defeito foi preenchido pelo coágulo e irradiado. Nos animais dos grupos do biomaterial e LED + biomaterial, os defeitos foram preenchidos com biomaterial e o último foi adicionalmente irradiado (λ=850±10 nm, 150 mW, Φ ~ 0.5 cm2, 20 J/cm2 - session, 140 J/cm2 -tratamento) a cada 48 h por duas semanas. Após a morte dos animais, amostras foram colhidas e analisadas por microscopia de luz, usando o grau de maturação do osso como marcador (deposição/organização das linhas basofílicas) no tecido ósseo neoformado. O grupo de LED + biomaterial apresentou processo de reparação mais avançado ao fim do experimento. Pode-se concluir que o uso da fototerapia LED foi eficaz na modulação positiva do processo de reparo ósseo de defeitos ósseos no fêmur de ratos submetidos ou não a enxerto com biomaterial.

Role of tricalcium phosphate implant in bridging the large osteoperiosteal gaps in rabbits.

Treatment options for large osteoperiosteal defects are limited and that which are available are not ideal. Osteoperiosteal defect were created in ulnae of both forelimbs of rabbits and tricalcium phosphate implant was used to bridge the gap. Amongst the 35 implanted ulnae, one implant got dislodged. Rest of the implants showed good adherence to host bone until the final follow up. Five control rabbit limbs (in which no implants were put) showed persistent bone gap. Histological and Electron microscopic examination revealed bone tissues covering the surface of the implant and bridging the gap. New bone was formed in the pores also. Tricalcium phosphate implants showed new bone formation due to osteoconductive properties. They are biodegradable. It is suggested that tricalcium phosphate implants are viable treatment alternatives in management of large osteoperiosteal defects with minimal to no adverse effects.

Effect of bleaching on sound enamel and with early artificial caries lesions using confocal laser microscopy

The aim of this study was to evaluate effect of bleaching agents on sound enamel (SE) and enamel with early artificial caries lesions (CL) using confocal laser scanning microscopy (CLSM). Eighty blocks (4 x 5 x 5 mm) of bovine enamel were used and half of them were submitted to a pH cycling model to induce CL. Eight experimental groups were obtained from the treatments and mineralization level of the enamel (SE or CL) (n=10). SE groups: G1 - unbleached (control); G2 - 4% hydrogen peroxide (4 HP); G3 - 4 HP containing 0.05% Ca (Ca); G4 - 7.5% hydrogen peroxide (7.5 HP) containing amorphous calcium phosphate (ACP). CL groups: G5 - unbleached; G6 - 4 HP; G7 - 4 HP containing Ca; G8 - 7.5 HP ACP. G2, G3, G6, G7 were treated with the bleaching agents for 8 h/day during 14 days, while G4 and G8 were exposed to the bleaching agents for 30 min twice a day during 14 days. The enamel blocks were stained with 0.1 mM rhodamine B solution and the demineralization was quantified using fluorescence intensity detected by CLSM. Data were analyzed using ANOVA and Fisher’s tests (α=0.05). For the SE groups, the bleaching treatments increased significantly the demineralization area when compared with the unbleached group. In the CL groups, no statistically significant difference was observed (p>0.05).The addition of ACP or Ca in the composition of the whitening products did not overcome the effects caused by bleaching treatments on SE and neither was able to promote remineralization of CL.

O objetivo deste estudo foi avaliar o efeito de agentes clareadores no esmalte sadio (ES) ou esmalte com lesão inicial de cárie artificial (LC), utilizando microscopia confocal laser de varredura (CLSM). Oitenta blocos (4 x 5 x 5 mm) de esmalte bovino foram usados, sendo que 40 destes foram desmineralizados com ciclagem de pH para induzir a LC. Oito grupos experimentais foram obtidos a partir dos tratamentos e condição do esmalte (ES ou LC), com n=10: Grupos ES: G1 - sem tratamento (controle); G2 - peróxido de hidrogênio 4% (PH4); G3: PH4 contendo 0,05% cálcio (Ca); G4 - peróxido de hidrogênio 7,5% (pH 7,5) contendo fosfato de cálcio amorfo (ACP). Grupos LC: G5 - não clareado; G6 - pH 4; G7 - pH 4 Ca; G8 - pH 7,5 ACP. Os grupos G2, G3, G6 e G7 foram tratados com o gel clareador por 8 h/dia durante 14 dias, enquanto as amostras dos grupos G4 e G8 foram submetidas ao agente clareador por 30 min/duas vezes ao dia, durante 14 dias. Os blocos de esmalte foram corados com solução de Rodamina B e a área fluorescente de desmineralização foi quantificada utilizando CLSM. Os dados foram submetidos a ANOVA e teste de Fisher (p<0,05). Para ES, os tratamentos clareadores aumentaram significativamente a área de desmineralização quando comparado com os grupos não clareados, entretanto, para LC não foi observado diferença estatística significante entre os grupos. A adição de ACP e Ca na composição dos géis clareadores não anulou os efeitos dos tratamentos clareadores no ES, assim como não teve capacidade de remineralizar o LC.

Enamel remineralization: controlling the caries disease or treating early caries lesions?

The emphasis currently given to new technologies for enamel remineralization suggests that the changes in the understanding of the dental caries disease, which occurred in the last century, were either not yet adopted or were forgotten. Just like in the past, when the disease was "treated" by restoring cavities, there is presently a misunderstanding on the concept of incipient lesion remineralization. The aim of this paper was to review some concepts about caries, the natural phenomenon of enamel remineralization and the effect of fluoride (F) on it, and also to discuss the clinical relevance of remineralizing products recently launched in the marketplace aiming to "treat early caries lesions".

Effects of Polycaprolactone-Tricalcium Phosphate, Recombinant Human Bone Morphogenetic Protein-2 and Dog Mesenchymal Stem Cells on Bone Formation: Pilot Study in Dogs

PURPOSE: The aim of this study was to evaluate the survival, proliferation, and bone formation of dog mesenchymal stem cells (dMSCs) in the graft material by using Polycaprolactone-tricalcium phosphate (PCL-TCP), auto-fibrin glue (AFG), recombinant human bone morphogenetic protein-2 (rhBMP-2), and dMSCs after a transplantation to the scapula of adult beagle dogs. MATERIALS AND METHODS: The subjects were two beagle dogs. Total dose of rhBMP-2 on each block was 10 microg with 50 microg/mg concentration. The cortical bone of the scapula of the dog was removed which was the same size of PCL-TCP block (Osteopore International Pte, Singapore; 5.0x5.0x8.0 mm in size), and the following graft material then was fixed with orthodontic mini-implant, Dual-top(R) (Titanium alloy, Jeil Co. Seoul, Korea). Four experimental groups were prepared for this study, Group 1: PCL-TCP + aFG; Group 2: PCL-TCP + aFG + dMSCs; Group 3: PCL-TCP + aFG + dMSCs + rhBMP-2; Group 4: PCL-TCP + aFG + dMSCs + rhBMP-2 + PCL membrane. The survival or proliferation of dMSCs cells was identified with an extracted tissue through a fluorescence microscope, H-E staining and Von-Kossa staining in two weeks and four weeks after the transplantation. RESULTS: The survival and proliferation of dMSCs were identified through a fluorescence microscope from both Group 1 and Group 2 in two weeks and four weeks after the transplantation. Histological observation also found that the injected cells were proliferating well in the G2, G3, and G4 scaffolds. CONCLUSION: This study concluded that bone ingrowth occurred in PCL-TCP scaffold which was transplanted with rhBMP-2, and MSCs did not affect bone growth. More sufficient healing time would be needed to recognize effects of dMSCs on bone formation.

An injectable cement: synthesis, physical properties and scaffold for bone repair.

Micro-invasive bone grafting is to deliver bone graft materials to the desired site through local puncturation and injection. It has many advantages such as little injury, simple procedures and high efficiency of osteogenesis. Limited sources of graft materials and complicated procedures are the main factors affecting the development of the technique. Therefore, to prepare a stable, economical, efficient and easy-to-use liquid graft material is crucial for the development of the technique. PURPOSE: The potential efficacy of an injectable calcium phosphate cement's handling properties and scaffold for bone repair performance was evaluated in a rabbit femoral condyles model. STUDY DESIGN: A comparative study was conducted between a new cement and a commercially available calcium phosphate cement. MATERIALS AND METHODS: The new cement and commercially available calcium-phosphate cements were compared in terms of the setting time, injection pressure, particle size and compressive strength. Then the cements were delivered to rabbit femoral condyles through local injection and then degradation of cements and bone formation were observed regularly after operation. RESULTS: The new injectable cement was superior to currently used cements in terms of permitted manipulation time, injection pressure, particle size, postoperative degradation and efficacy of scaffold for bone repair; nevertheless, the new cement was slightly inferior to currently used cements in compressive strength and the final setting time. CONCLUSIONS: The new injectable cement is more suitable for the clinical study of micro-invasive bone grafting. It allows a brand new bone grafting procedure and provides a new graft material and thus merits further development and wider application.

Comparative therapeutic use of Risedronate and Calcarea phosphorica - allopathy versus homeopathy - in bone repair in castrated rats / Comparação do uso terapêutico de Risedronato e Calcarea phosphorica - alopatia versus homeopatia - no reparo ósseo em ratos castrados

Osteoporosis, a disease characterized by progressive bone loss, has been the target of several studies in the past few years. It results in a much higher risk for fractures and might cause slower bone lesion healing. The aim of this work was to study the effects of Risedronate (allopathic medicine) and Calcarea phosphorica 6CH (homeopathic medicine) on the repair of bone lesions in male rats with osteoporosis induced by castration. Eighty-four three-month-old rats were used divided into four groups of twenty-one animals each. Three groups where castrated and one group was submitted to Sham surgery. One month later, cortical lesions were made in all animals' tibiae and, after one day, the different experimental treatments began according to the following groups: CR - castrated/Risedronate (1 mg/kg/day); CCp - castrated/Calcarea phosphorica 6CH (3 drops/day); CP - castrated/placebo and SP - Sham/placebo. The animals were sacrificed at seven, fourteen and twenty-eight days after the beginning of the treatments and had their tibiae removed. Digital radiographs of the tibiae were taken and analyzed in order to evaluate the optical density of the defect area. Then, they were decalcified and processed for histological and histomorphometrical analysis. The data were submitted to ANOVA, and to the Tukey and Dunnett tests (5 percent). The allopathic and homeopathic treatments led to different bone formation as regards remodeling and maturation aspects. Further research is necessary to access the resistance and quality of the newly formed bone.

A osteoporose, doença caracterizada pela perda de massa óssea, tem sido alvo de estudos nos últimos anos. Fraturas decorrentes da osteoporose são muito comuns e podem apresentar consolidação mais lenta. O objetivo deste trabalho foi avaliar o efeito do risedronato (medicamento alopático) e da Calcarea phosphorica 6CH (medicamento homeopático) no reparo de lesões ósseas em ratos com osteoporose induzida por castração. Para tanto, foram utilizados oitenta e quatro ratos, com três meses de idade, separados em quatro grupos de vinte e um animais, sendo três grupos submetidos à castração e um grupo a falsa cirurgia ("Sham"). Um mês após a cirurgia, foram realizadas lesões corticais na tíbia de todos os animais e, a partir do dia seguinte, os tratamentos experimentais foram iniciados de acordo com os seguintes grupos: CR - castrado/risedronato (1 mg/kg/dia); CCp - castrado/Calcarea phosphorica 6CH (três gotas/dia); CP - castrado/placebo e SP - "sham"/placebo. Os animais foram sacrificados aos sete, catorze e vinte e oito dias após o início do tratamento e as tíbias foram removidas. Radiografias digitais foram realizadas e avaliadas para obter a densidade óptica na área do defeito. Em seguida, foram descalcificadas e processadas para análise histológica e histomorfométrica. Os dados foram submetidos a ANOVA e aos testes de Tukey e Dunnett (5 por cento). Os tratamentos alopático e homeopático levaram a formação óssea de aspecto diferente considerando a remodelação e maturação. Mais pesquisas são necessárias para avaliar a resistência e qualidade do osso neoformado.

Guided bone regeneration with beta-tricalcium phosphate and poly Llactide-co-glycolide-co-epsilon-caprolactone membrane in partial defects of canine humerus

This study was performed to evaluate the effect of betatricalcium phosphate and poly L-lactide-co-glycolide-coepsilon- caprolactone (TCP/PLGC) membrane in the repair of partial bone defects in canine proximal humerus. Three adult mixed-breed dogs were used during the experimental period. The length of the defect was quarter of the full length of humerus, and width of the defect was quarter of middle diameter of the lateral aspect of humerus. The humeri of each dog were divided into treatment (TCP/ PLGC) and control groups. The defect was covered with TCP/PLGC membrane in treatment group. To evaluate regeneration of the bone, computerized tomography (CT) and histopathologic examination were performed. The radiopaque lines were appeared at the original defect sites in TCP/PLGC group but below the original site in control at 4th week. Radiopacity and thickness of the defect sites, and radiopaque lines were more increased at 8th week than those of 4th week. Histopathologic findings revealed fibrous connective tissue migration into the defect and the migration inhibited the structure of new cortex to be placed in the original level in control whereas new cortex growth was found in the level of original line in TCP/ PLGC group. However, the new cortical bone in the TCP/ PLGC group was thinner and less organized than the adjacent intact cortex, and the amount of new cancellous bones were also scanty. The result suggested that TCP/ PLGC membrane is a good guided bone regeneration material to restore the original morphology of humerus in partial defect.

Repair of sheep metatarsus defects by using tissue-engineering technique / 华中科技大学学报（医学）（英德文版）

Tissue-engineering bone with porous ,betatricalcium phosphate (3-TCP) ceramic and autologous bone marrow mesenchymal stem cells (MSC) was constructed and the effect of this composite on healing of segmental bone defects was investigated. 10-15 ml bone marrow aspirates were harvested from the iliac crest of sheep, and enriched for MSC by density gradient centrifugation over a Percoll cushion (1. 073 g/ml). After cultured and proliferated, tissue-engineering bones were constructed with these,cellS seeded onto porous f-TCP, and then the constructs were implanted in 8 sheep left metatarsus defect (25 mm in length) as experimental group. Porous ,-TCP only were implanted to bridge same size and position defects in 8 sheep as control group, and 25 mm segmental bone defects of left metatarsus were left empty in 4 sheep as blank group. Sheep were sacrificed on the 6th, 12th, and 24th week postoperatively and the implants samples were examined by radiograph, histology, and biomechanical test. The 4 sheep in blank group were sacrificed on the 24th week postoperatively. The results showed that new bone tissues were observed either radiographic or histologically at the defects of experimental group as early as 6th week postoperatively, but not in control group, and osteoid tissue, woven bone and lamellar bone occurred earlier than in control group in which the bone defects were repaired in "creep substitution" way, because of the new bone formed in direct manner without progression through a cartilaginous intermediate. At the 24th week, radiographs and biomechanical test revealed an almost complete repair of the defect of experimental group, only partly in control group. The bone defects in blank group were non-healing at the 24th week. It was concluded that engineering bones constructed with porous -TCP and autologous MSC were capable of repairing segmental bone defects in sheep metatarsus beyond "creep substitution" way and making it healed earlier. Porous ,-TCP being constituted with autologous MSC may be a good option in healing critical segmental bone defects in clinical practice and provide insight for future clinical repair of segmental defect.

Effect of calcium phosphate ceramic implants used for mandibular ridge augmentation on gingival cells. an ultrastructural study

Six adult dogs were injected with calcium phosphate ceramic material through subperiosteal tunneling technique. Gingival specimens were taken after 4, 8 and 12 months from both sides of the mandible, the injected side [study specimens] and the opposite side [control specimens] to be processed for light and electron microscopy. The implant particles were detected in gingiva after 4 and 8 months. After 12 months they were detected only on ultrastructural level. In epidermis, implanted particles appeared as dense granules in membrane bound vacuoles in perinuclear zone. No epidermal reaction was noticed, except for increased cellular attachment. In dermis the particles were phagocytosed inside macrophages. Mild activation of fibroblasts, plasma cells, lymphocytes and leukocytes was noticed with mild collagen deposition