Late hyaluronidase injection in local anesthesia: Morphofunctional evaluation in rat sciatic nerve block

Introduction: Despite the enhancing effects of hyaluronidase (HYAL) over duration of anesthesia, this enzyme could cause adverse effects when injected concomitantly with local anesthetics in dental blocks. Objective: This study aimed to assess the tissue alterations caused by a local anesthetic protocol consisting of a late HYAL injection and confirm its functional effectiveness. Materials and Methods: The protocol efficacy was proved by evaluating sensory and motor functions in rats. The sciatic nerve was blocked with 2% lidocaine (LID) with epinephrine (n = 25). Thirty minutes later, 75 TRU/ml HYAL was injected into the same site (experimental group, LID/HYAL). One week later, this protocol was repeated in the contralateral hindlimb, injecting only HYAL's vehicle (control group, LID/vehicle [LID/V]). To observe the integrity of the local tissues, histological specimens were obtained 1, 24, 48, and 72 h after treatment with LID/HYAL or LID/V (n = 16 each) and stained with hematoxylin/eosin and picrosirius red. Results: Local inflammation was similar in both groups. The integrity of the nerve fibers was preserved, in spite of some inflammation-associated injuries in the surrounding tissues. The reversible tissue disorganization caused by HYAL, probably facilitated the diffusion of the residual anesthetic to the nerve, resulting in a prolonged anesthetic effect (P < 0.05). Conclusions: No irreversible morphological alterations are caused by the administration of HYAL prior the end of the LID-induced block. Moreover, this protocol prolongs LID's anesthetic effect.

Effects of low molecular weight heparin on alveolar bone loss in wistar rats

Abstract This study aimed to assess the effects of low molecular weight heparin (LMWH) on alveolar bone loss (ABL), blood count, and counting of megakaryocytes and adipocytes in male Wistar rats. Forty male 60-day Wistar rats were randomly divided into four groups: Control (C), Periodontal Disease (PD), Heparin (Hp) and Heparin + Periodontal Disease (Hp+PD). LMWH was applied for 60 days at doses of 1 ml/kg/day. Blood samples were collected at baseline, 30 and 60. On day-49, PD and Hp+PD groups were subjected to ligature-induced periodontitis around second upper right molar. The left side was assessed as spontaneous alveolar bone loss. Mean ABL in the side with ligature showed significantly different between C (0.35±0.07 mm) and Hp+DP (0.49±0.09 mm) groups (p<0.001), between PD (0.55±0.11 mm) and Hp (0.32±0.06 mm) groups (p<0.001) and between Hp and Hp+DP groups (p<0.001). No significant differences were found among groups for ABL in the side without ligature. Animal weight, food intake, and water consumption showed no statistically significant difference among groups. Megakaryocytes and adipocytes were counted using optical microscopy and no statistically significant differences were found. Within-groups, there were an increase and a decrease, respectively, in the counting of lymphocytes (p=0.005 for C and p=0.009 for Hp+PD groups only) and leukocytes (p=0.003 for C, p=0.001 for PD, p=0.002 for Hp, and p<0.001 for Hp+PD groups). There was no decrease in the number of platelets in the three collection periods. LMWH was not able to affect ABL, but it may change the blood counting, especially increasing lymphocytes.

Resumo O presente estudo objetivou verificar o efeito da heparina de baixo peso molecular (HBPM) sob a perda óssea alveolar (POA), contagem de células sanguíneas, megacariócitos e adipócitos em ratos Wistar machos. Quarenta ratos Wistar de 60 dias foram randomicamente divididos em quatro grupo: Controle (C), Doença Periodontal (DP), Heparina (Hp) e Heparina + Doença Periodontal (Hp+DP). HBPM foi aplicada durante 60 dias em doses de 1 mL/kg/dia. Coletas sanguíneas foram realizadas nos dias 0, 30 e 60. No dia 49, os grupos DP e Hp+DP receberam indução de doença periodontal por ligadura ao redor do segundo molar superior direito. No lado esquerdo, verificou-se perda óssea alveolar espontânea. A média de POA no lado com ligadura mostrou-se estatisticamente diferente entre os grupos C (0,35±0,07 mm) e Hp+PD (0,49±0,09 mm) (p<0,001), entre os grupos DP (0,55±0,11 mm) e Hp (0,32±0,06 mm) (p<0,001) e entre os grupos Hp e Hp+DP (p<0,001). Nenhuma diferente significativa foi observada entre os grupos no lado sem ligadura. Peso dos animais, consumo de ração e ingestão de água não mostraram diferenças significativas entre os grupos. Megacariócitos e adipócitos foram contados por microscopia óptica e nenhuma diferença significativa foi encontrada. Dentro dos grupos, houve um aumento e uma diminuição, respectivamente, na contagem de linfócitos (p=0,005 no grupo C e p=0,009 no grupo Hp+DP somente) e leucócitos (p=0,003 no grupo C, p=0.001 no grupo DP e p=0,002 no grupo Hp e Hp+DP). Não houve diminuição no número de plaquetas nos três períodos de coleta. HBPM não foi capaz de modificar a POA, porém modificou a contagem de células sanguíneas, especialmente aumentando o número de linfócitos.

Effects of diabetes and hypertension on oral mucosa and tgfß1 salivary levels

Abstract The aim of this study was to investigate salivary levels of TGFβ1 and proliferation/ maturation of epithelial mucosa cells in diabetic and hypertensive patients. Design: in this cross-sectional study, whole stimulated saliva and oral mucosa exfoliative cytology specimens were collected from 39 patients that were healthy (control, n=10) or presented history of arterial hypertension (HAS, n=9), diabetes mellitus (DM, n=10) or both (DM+HAS, n=10). Salivary flow rate (SFR), TGFβ1 level in saliva, AgNORs and the epithelial maturation were evaluated. Non-parametric Kruskal-Wallis test, followed by Dunn's multiple comparison post-test and the Spearman test correlation analysis were used. SFR showed a significant decreased in DM and DM+HAS (0.47±0.11 and 0.64±0.43 mL/min) when compared to control (1.4±0.38 mL/min). DM+HAS presented the highest value of TGFβ1 concentration (24.72±5.89 pg/mL). It was observed a positive correlation between TGFβ1 and glycaemia (R=0.6371; p<0.001) and a negative correlation between TGFβ1 and saliva (R=-0.6162; p<0.001) and glycaemia and SFR (R=-0.5654; P=0.001). AgNORs number and status of maturation of mucosa cells were similar for all conditions. DM and DM+HAS presented the lowest SFR, which correlated with increased TGFβ1 levels. Despite the higher TGFβ1 secretion it was not observed changes in the morphology or proliferation of epithelial cells when diabetes or hypertension was present.

Resumo O objetivo deste estudo foi investigar os níveis de TGFβ1 na saliva e a proliferação/maturação das células epiteliais da mucosa em paciente diabéticos e hipertensos. Neste estudo transversal, saliva estimulada e amostras de citologia exfoliativa de mucosa oral foram coletadas de um total de 39 pacientes que se apresentavam saudáveis (controle, n=10) ou com história de hipertensão arterial (HAS, n=9), diabetes mellitus (DM, n=10) ou ambos (DM+HAS, n=10). Taxa de fluxo salivar (SFR), níveis de TGFβ1 na saliva, AgNORs e maturação epitelial foram avaliados. Teste não-paramétrico de Kruskal-Wallis, seguido de comparação múltipla de Dunn e correlação de Spearman foram utilizados para as análises. SFR diminuiu significantemente em DM e DM+HAS (0,47±0,11 e 0,64±0,43 mL/min) quando comparado ao controle (1,4±0,38 mL/min). DM+HAS apresentou os maiores valores de concentração de TGFβ1 (24,72±5,89 pg/mL). Foi observada uma correlação positiva entre TGFβ1 e glicemia (R=0,6371; p<0,001) e uma correlação negativa entre TGFβ1 e saliva (R=-0,6162; p<0,001) e glicemia e SFR (R=-0,5654; p=0,001). Número de AgNORs e o padrão da maturação das células epiteliais foram similares entre os todos grupos. DM e DM+HAS apresentaram os menores valores de SFR, os quais foram correlacionados com o aumento nos níveis de TGFβ1. Apesar da maior secreção de TGFβ1, não foram observadas mudanças na morfologia ou proliferação das células epiteliais quando o paciente apresentava diabetes ou hipertensão.