Histometric study of alveolar bone healing in rats treated with the nonsteroidal anti-inflammatory drug nimesulide.

PURPOSE: There is extensive experimental and clinical evidence in the orthopedic area that prolonged use of nonselective (inhibitor of both cyclooxygenases 1 and 2) nonsteroidal anti-inflammatory drugs can hinder long bone fracture healing, spinal fusion rate, and new bone formation around implants. The purpose of the present study was to investigate whether nimesulide (Nimesulida, Medley S.A., Campinas, SP, Brazil), a preferential cyclooxygenase-2 inhibitor, can hinder alveolar bone healing, in rats. MATERIALS AND METHODS: Treated rats received oral doses (5 mg/kg/rat/day) of nimesulide from the day of tooth extraction until euthanasia 2 weeks later and control rats received tap water (n = 5 per group). The volume of neoformed bone inside the alveolar socket was estimated in semiserial longitudinal histological sections by a differential point-counting method, and the significance of the difference between groups was analyzed by Student t test (P < 0.05 for statistical significance). RESULTS: Histometric data confirmed histological observation that the volume fraction of new bone trabeculae in treated rats was not significantly different from that in control rats. CONCLUSION: Short-term treatment with nimesulide, although its capacity to inhibit preferentially the enzyme cyclooxygenase-2, does not interfere with alveolar bone healing in rats.

Treatment with paracetamol, ketorolac or etoricoxib did not hinder alveolar bone healing: a histometric study in rats

Prostaglandins control osteoblastic and osteoclastic function under physiological or pathological conditions and are important modulators of the bone healing process. The non-steroidal anti-inflammatory drugs (NSAIDs) inhibit cyclooxygenase (COX) activity and consequently prostaglandins synthesis. Experimental and clinical evidence has indicated a risk for reparative bone formation related to the use of non-selective (COX-1 and COX-2) and COX-2 selective NSAIDs. Ketorolac is a non-selective NSAID which, at low doses, has a preferential COX-1 inhibitory effect and etoricoxib is a new selective COX-2 inhibitor. Although literature data have suggested that ketorolac can interfere negatively with long bone fracture healing, there seems to be no study associating etoricoxib with reparative bone formation. Paracetamol/acetaminophen, one of the first choices for pain control in clinical dentistry, has been considered a weak anti-inflammatory drug, although supposedly capable of inhibiting COX-2 activity in inflammatory sites. OBJECTIVE: The purpose of the present study was to investigate whether paracetamol, ketorolac and etoricoxib can hinder alveolar bone formation, taking the filling of rat extraction socket with newly formed bone as experimental model. MATERIAL AND METHODS: The degree of new bone formation inside the alveolar socket was estimated two weeks after tooth extraction by a differential point-counting method, using an optical microscopy with a digital camera for image capture and histometry software. Differences between groups were analyzed by ANOVA after confirming a normal distribution of sample data. RESULTS AND CONCLUSIONS: Histometric results confirmed that none of the tested drugs had a detrimental effect in the volume fraction of bone trabeculae formed inside the alveolar socket.

Cola beverage consumption delays alveolar bone healing: a histometric study in rats.

Epidemiological studies have suggested that cola beverage consumption may affect bone metabolism and increase bone fracture risk. Experimental evidence linking cola beverage consumption to deleterious effects on bone is lacking. Herein, we investigated whether cola beverage consumption from weaning to early puberty delays the rate of reparative bone formation inside the socket of an extracted tooth in rats. Twenty male Wistar rats received cola beverage (cola group) or tap water (control group) ad libitum from the age of 23 days until tooth extraction at 42 days and euthanasia 2 and 3 weeks later. The neoformed bone volume inside the alveolar socket was estimated in semi-serial longitudinal sections using a quantitative differential point-counting method. Histological examination suggested a decrease in the osteogenic process within the tooth sockets of rats from both cola groups, which had thinner and sparser new bone trabeculae. Histometric data confirmed that alveolar bone healing was significantly delayed in cola-fed rats at three weeks after tooth extraction (ANOVA, p = 0.0006, followed by Tukey's test, p < 0.01). Although the results of studies in rats cannot be extrapolated directly to human clinical dentistry, the present study provides evidence that cola beverage consumption negatively affect maxillary bone formation.

Cola beverage consumption delays alveolar bone healing: a histometric study in rats

Epidemiological studies have suggested that cola beverage consumption may affect bone metabolism and increase bone fracture risk. Experimental evidence linking cola beverage consumption to deleterious effects on bone is lacking. Herein, we investigated whether cola beverage consumption from weaning to early puberty delays the rate of reparative bone formation inside the socket of an extracted tooth in rats. Twenty male Wistar rats received cola beverage (cola group) or tap water (control group) ad libitum from the age of 23 days until tooth extraction at 42 days and euthanasia 2 and 3 weeks later. The neoformed bone volume inside the alveolar socket was estimated in semi-serial longitudinal sections using a quantitative differential point-counting method. Histological examination suggested a decrease in the osteogenic process within the tooth sockets of rats from both cola groups, which had thinner and sparser new bone trabeculae. Histometric data confirmed that alveolar bone healing was significantly delayed in cola-fed rats at three weeks after tooth extraction (ANOVA, p = 0.0006, followed by Tukey's test, p < 0.01). Although the results of studies in rats cannot be extrapolated directly to human clinical dentistry, the present study provides evidence that cola beverage consumption negatively affect maxillary bone formation.

Treatment with paracetamol, ketorolac or etoricoxib did not hinder alveolar bone healing: a histometric study in rats.

UNLABELLED: Prostaglandins control osteoblastic and osteoclastic function under physiological or pathological conditions and are important modulators of the bone healing process. The non-steroidal anti-inflammatory drugs (NSAIDs) inhibit cyclooxygenase (COX) activity and consequently prostaglandins synthesis. Experimental and clinical evidence has indicated a risk for reparative bone formation related to the use of non-selective (COX-1 and COX-2) and COX-2 selective NSAIDs. Ketorolac is a non-selective NSAID which, at low doses, has a preferential COX-1 inhibitory effect and etoricoxib is a new selective COX-2 inhibitor. Although literature data have suggested that ketorolac can interfere negatively with long bone fracture healing, there seems to be no study associating etoricoxib with reparative bone formation. Paracetamol/acetaminophen, one of the first choices for pain control in clinical dentistry, has been considered a weak anti-inflammatory drug, although supposedly capable of inhibiting COX-2 activity in inflammatory sites. OBJECTIVE: The purpose of the present study was to investigate whether paracetamol, ketorolac and etoricoxib can hinder alveolar bone formation, taking the filling of rat extraction socket with newly formed bone as experimental model. MATERIAL AND METHODS: The degree of new bone formation inside the alveolar socket was estimated two weeks after tooth extraction by a differential point-counting method, using an optical microscopy with a digital camera for image capture and histometry software. Differences between groups were analyzed by ANOVA after confirming a normal distribution of sample data. RESULTS AND CONCLUSIONS: Histometric results confirmed that none of the tested drugs had a detrimental effect in the volume fraction of bone trabeculae formed inside the alveolar socket.

Comparison of rat bone healing following intra-alveolar grafting with organic or inorganic bovine bone particles

Aim: This study compared, histometrically, the alveolar bone healing after grafting rats extraction socket with particles of organic or inorganic bovine bone. Method: The volume fraction of grafted materials and bone trabeculae was estimated in histologic images at the end of the 2nd and 9th weeks post-operatively by a differential point-counting method. Results: Particles of both materials were observed partially filling the cervical alveolar third and the volume fraction of inorganic graft was larger than that of organic graft 2 and 9 weeks following implantation. Although evoking neither a foreign-body reaction nor a persisting inflammatory response, both materials delayed bone healing. By the 2nd week, the delay was more pronounced in the animals grafted with inorganic than in those grafted with organic bone, but only in the animals whose inorganic graft occupied more than 50% of the cervical third. By the 9th week, despite the greater volume fraction of inorganic graft the percent of bone healing was similar to that observed in the animals grafted with organic bone. Conclusion: The degree of impairment of bone healing resulted from combination of factors such as type of material, its relative amount and the phase of the reparational process.

Grafting of tooth extraction socket with inorganic bovine bone or bioactive glass particles: comparative histometric study in rats.

PURPOSE: To compare histometrically, in rats, the bone healing after grafting the incisor extraction sockets with inorganic bovine bone or bioactive glass particles. MATERIAL: The volume fraction of grafted materials and alveolar healing components was estimated in histologic images at the end of the second and ninth weeks postoperatively by a differential point-counting method. RESULTS: Both materials were histologically observed partially filling the cervical alveolar third and, although evoking neither a foreign body reaction nor a persisting inflammatory response, delayed new bone formation in trial areas around their particles. By the second week, the delay in bone healing was more pronounced in the animals grafted with inorganic than in those grafted with bioactive glass, and an opposing result was observed during a 9-week period. CONCLUSION: Both inorganic bovine bone and bioactive glass particles grafted in the incisor extraction sockets of rats delayed new bone formation, and the degree of impairment resulted from a combination of factors such as type of material and phase of the reparation process.

Alveolar wound healing after implantation with a pool of commercially available bovine bone morphogenetic proteins (BMPs): a histometric study in rats.

The capacity of a commercially available pool of bovine bone morphogenetic proteins (BMPs) to stimulate osteogenesis in the rat alveolar healing was investigated by histometric analysis. Male rats were anesthetized and had their upper incisor extracted. A pool of purified bovine BMPs adsorbed to microgranular resorbable hydroxyapatite was agglutinated with bovine collagen and saline before implantation into the alveolar socket. The implanted and control rats (n=30 per group) were sacrificed 1 to 9 weeks postoperatively, the hemi-maxillae were decalcified, processed for paraffin embedding and semi-serial longitudinal sections were obtained and stained with hematoxylin and eosin. The volume fraction of alveolar healing components was estimated by a differential point-counting method in histologic images. The results showed that in both, control and implanted rats, the alveolar healing followed the histologic pattern usually described in the literature. Quantitative data confirmed that the BMPs mixture did not stimulate new bone formation in the alveolar socket of implanted rats. These results suggest that the pool of BMPs adsorbed to hydroxyapatite and agglutinated with bovine collagen did not warrant incorporation of the osteoinductive proteins to a slow-absorption system that would allow a BMPs release rate compatible to that of new bone formation, and thus more adequate to osteoinduction.

Diagnóstico e tratamento de fibroma ossificante periférico gengival: relato de caso / Diagnosis and treatment of gingival peripheral ossifying fibroma: a case report

O fibroma ossificante periférico é uma lesão de natureza não-neoplásica proliferativa, sendo normalmente assintomática e de evolução lenta. Apresenta características clínicas similares a outros processos não-neoplásicos, sendo necessário um diagnóstico diferencial entre os diferentes tipos de lesões que normalmente acometem a gengiva (granuloma piogênico, fibroma de células gigantes, granuloma periférico de células gigantes e fibroma de irritação). O objetivo deste trabalho é relatar um caso clínico de fibroma ossificante periférico gengival, enfocando-se para o diagnóstico e tratamento.

The peripheral ossifying fibroma (POF) is an asymptomatic non-neoplasic proliferating process with low evolution. The POF shows similar clinical characteristics to the others gingival lesions (pyogenic granuloma, giant cell fibroma, peripheral giant cell granuloma and irritation fibroma). The purpose of this study is to report a clinical case of gingival peripheral ossifying fibroma, focusing for the diagnosis and treatment.

Alveolar wound healing after implantation with a pool of commercially available bovine bone morphogenetic proteins (BMPs): a histometric study in rats

The capacity of a commercially available pool of bovine bone morphogenetic proteins (BMPs) to stimulate osteogenesis in the rat alveolar healing was investigated by histometric analysis. Male rats were anesthetized and had their upper incisor extracted. A pool of purified bovine BMPs adsorbed to microgranular resorbable hydroxyapatite was agglutinated with bovine collagen and saline before implantation into the alveolar socket. The implanted and control rats (n=30 per group) were sacrificed 1 to 9 weeks postoperatively, the hemi-maxillae were decalcified, processed for paraffin embedding and semi-serial longitudinal sections were obtained and stained with hematoxylin and eosin. The volume fraction of alveolar healing components was estimated by a differential point-counting method in histologic images. The results showed that in both, control and implanted rats, the alveolar healing followed the histologic pattern usually described in the literature. Quantitative data confirmed that the BMPs mixture did not stimulate new bone formation in the alveolar socket of implanted rats. These results suggest that the pool of BMPs adsorbed to hydroxyapatite and agglutinated with bovine collagen did not warrant incorporation of the osteoinductive proteins to a slow-absorption system that would allow a BMPs release rate compatible to that of new bone formation, and thus more adequate to osteoinduction.

No presente trabalho analisou-se histometricamente a capacidade de um material de origem nacional em estimular o reparo ósseo alveolar de ratos. Uma mistura de BMPs bovinas purificadas adsorvidas à hidroxiapatita microgranular absorvível foi aglutinada com colágeno bovino e soro fisiológico e implantada na cavidade de extração de incisivos superiores. Os ratos implantados e controles (n=30 por grupo) foram sacrificados após 1, 2, 3 e 9 semanas, as hemi-maxilas contendo os alvéolos em reparação foram descalcificadas e processadas para inclusão em parafina e obtenção de cortes semi-seriados, corados com hematoxilina e eosina. O volume percentual de tecido ósseo reparacional foi estimado por um método de contagem diferencial de pontos, em imagens histológicas analisadas sob uma ocular contendo um retículo com 100 pontos eqüidistantes. Nos ratos controles e implantados o reparo seguiu o padrão descrito na literatura. No grupo implantado, o material não estimulou o reparo ósseo alveolar em nenhum dos períodos analisados. Os resultados sugerem que a mistura de BMPs adsorvidas à hidroxiapatita microgranular absorvível não tenha garantido sua incorporação a um sistema carreador de absorção lenta que propiciasse sua liberação num ritmo compatível com o da neoformação óssea, portanto, adequado à osteoindução.

Bone healing in osteoporotic female rats following intra-alveolar grafting of bioactive glass.

We have investigated the effect of ovariectomy combined with a low Ca diet on bone healing following the implantation of bioactive glass into extraction sockets, in rats. Ovariectomized rats received a low Ca diet from the day of surgery until sacrifice while sham-operated animals were fed a standard laboratory chow. Two weeks after surgery the upper incisors were extracted and the alveolar sockets in both groups were partially filled with a particulate bioglass (PerioGlas). The animals were killed 1, 2, 3 and 9 weeks after tooth extraction and the relative volume fraction of the healing components (bone trabeculae, connective tissue and coagulum remnants) was estimated in histological paraffin sections by a histometric differential point-counting method. The bioglass particles persisted inside the socket for all the experimental periods and, as bone repair proceeded, they were progressively enclosed in newly formed bone trabeculae which in some cases established a close contact with their surface. The volume fraction of neoformed bone trabeculae relative to the volume fraction of connective tissue and coagulum remnants was greater in the sockets of ovariectomized animals implanted with bioglass than in those of the overiectomized non-implanted groups.

Comparison between two experimental protocols to promote osteoporosis in the maxilla and proximal tibia of female rats

Comparou-se o efeito de dois protocolos experimentais (ovariectomia associada ou não à dieta pobre em Ca++) utilizados para promover osteoporose em maxila e metáfise proximal de ratas, nos períodos de 5 e 11 semanas pós-cirurgia. Ratas Wistar foram ovariectomizadas ou submetidas à cirurgia simulada. Metade das ratas ovariectomizadas recebeu dieta pobre em Ca++ (ovx*) e as demais (ovx), assim como as que sofreram falsa cirurgia, receberam dieta comercial. Foram coletados o osso maxilar (após extração dos molares) e a metáfise proximal da tíbia para medidas do peso seco (60ºC/12 h) e do da cinza óssea (700ºC/14 h), utilizada para dosagem de Ca++ (método colorimétrico). Cinco semanas após a cirurgia, não se observaram alterações nos parâmetros investigados no grupo ovx, enquanto no grupo ovx* houve redução da massa óssea da metáfise proximal (17 por cento) e da maxila (35 por cento). Após 11 semanas, o grupo ovx apresentou 14 por cento de redução da massa óssea da metáfise proximal, mas não da maxila, enquanto no grupo ovx* observou-se diminuição de 30 por cento em ambos os segmentos ósseos. A concentração de Ca++ na cinza não se alterou em nenhuma condição experimental. Os resultados mostram que apenas a deficiência de estrógeno não é suficiente para provocar osteoporose maxilar em ratas num período de até 11 semanas, mas que nesse período já se observa seu efeito deletério na massa da metáfise proximal. Quando se associa a ovariectomia à dieta pobre em Ca++, observa-se diminuição da massa óssea após 5 semanas, 2 vezes maior na maxila do que na metáfise proximal da tíbia.

Comparison between two experimental protocols to promote osteoporosis in the maxilla and proximal tibia of female rats.

The effects of two experimental protocols (ovariectomy associated or not with a low calcium diet) used to promote osteoporosis in the rat maxilla and proximal tibia were compared 5 and 11 weeks after surgery. Female Wistar rats were ovariectomized or sham-operated. Half of the ovariectomized rats were fed a low Ca++ diet (ovx*) and the remaining ovariectomized (ovx) and sham animals received a standard chow. At sacrifice, the proximal metaphysis was excised from the tibia and the molars were extracted from the hemi-maxilla. Dry (60 degrees C overnight) and ash (700 degrees C/14 h) weights were measured and the ashes were used for Ca++ measurement by means of a colorimetric method. After 5 weeks, ovx caused no alteration while ovx* decreased proximal metaphysis (17%) and maxilla (35%) bone mass. After 11 weeks, ovx caused a 14% bone mass reduction in the proximal metaphysis but not in the maxilla, while ovx* caused a comparable bone mass reduction (30%) in both bone segments. Calcium concentration was not altered in any experimental condition. The results show that estrogen deficiency is insufficient to cause maxillary osteoporosis in rats over an 11-week period and a long-term ovariectomy is needed to exert deleterious effect on proximal metaphysis bone mass. When a low Ca++ diet is associated with estrogen deficiency, however, a relatively precocious harmful effect is observed, twice as pronounced in the maxilla than in the proximal metaphysis. On a long-term basis, ovariectomy associated with a low Ca++ diet seems to be equally injurious to both proximal metaphysis and maxilla.

Implante de um floculado de resina de mamona em alvéolo dental de rato / Implantation of flakes of castor oil resin in rat dental alveolus

Os objetivos do presente trabalho foram: 1) testar a biocompatibilidade de uma resina natural, derivada do óleo de mamona, implantada na cavidade de extraçäo dental de ratos, e 2) estudar a possível interferência do material na cronologia do reparo alveolar. O material (AUG-EX, Poliquil Araraquara Polímeros Químicos Ltda., Araraquara - SP) foi implantado no alvéolo imediatamente após a extraçäo do incisivo superior direito e os ratos foram sacrificados de 1 a 6 semanas após a extraçäo ou extraçäo + implante. As hemimaxilas foram descalcificadas e processadas para inclusäo em parafina e obtençäo de cortes semi-seriados, corados com hematoxilina-eosina. Os flocos da resina, de forma irregular e tamanho variável, localizaram-se entre os terços alveolares médio e cervical, inicialmente circundados por tecido de granulaçäo e a seguir por quantidade progressivamente maior de tecido ósseo, no geral com a presença de um tecido conjuntivo interposto, mas em algumas áreas estabelecendo aparente osseointegraçäo direta. Näo houve persistência da reaçäo inflamatória, mas observou-se pequena quantidade de células gigantes aderidas à superfície do material, em todos os períodos. A análise histométrica (contagem diferencial de ponto) do terço apical mostrou um atraso de 13 por cento a 20 por cento no reparo alveolar dos ratos implantados, com menor neoformaçäo óssea associada a maiores volumes percentuais de tecido conjuntivo e de remanescentes do coágulo sangüíneo