Viability of fibroblasts in a novel probiotic storage media

Abstract – A number of storage media have been investigated as to their ability to maintain the viability of the periodontal ligament (PDL) cells and thus to permit longer extra‐alveolar periods prior to replantation of avulsed teeth. The aim of the present in vitro study was to evaluate the number of viable PDL cells of avulsed teeth treated by Hank’s Balanced Salt Solutions (HBSS), saline, a novel probiotic solution and milk. Thirty‐six freshly extracted single‐rooted human teeth with closed apices were divided into one of the four experimental groups and two control groups (N = 6 each). The positive and negative controls corresponded to 0 min and an 8‐h dry time respectively. Following extraction, the coronal 3 mm of PDL tissue was scraped with a #15 scalpel to remove cells that might have been damaged. The experimental teeth were dried for 30 min followed by a 45 min immersion in one of the four experimental media. Each experimental tooth, after drying and soaking, was incubated for 30 min with a 2.5 ml solution of 0.2 mg ml^?1 of collagenase CLS II and a 2.4 mg ml^?1 solution of dispase grade II in phosphate buffer saline (PBS). The cells were then labelled with 0.4% Trypan blue for determination of viability. The teeth stored in positive control demonstrated the highest number of viable PDL cells followed in rank order by HBSS, saline, Lactobacillus reuteri solution and milk. There was no significant difference in the number of viable PDL cells between HBSS, milk, L. reuteri solution and saline. Within the parameters of this study, it appears that probiotic may be able to maintain PDL cell viability as HBSS, milk, or saline.     

Evaluation of the Toxicity and Reversibility Profile of the Aqueous Seed Extract of Hunteria Umbellata (K. Schum.) Hallier F. in Rodents

Hunteria umbellata (K. Schum.) Hallier f. (family: Apocynaceae) is reputed for the folkloric management of labour, pain and swellings, stomach ulcers, diabetes, obesity, and anaemia, with no scientific report of its toxicity and reversibility profile. The present study was, therefore, aimed at investigating the in vivo toxicity and reversibility profile of the aqueous seed extract of Hunteria umbellata (HU). The acute oral and intraperitoneal toxicity studies of HU were determined in Swiss albino mice while its 90-day oral toxicity and toxicity reversibility profile on anthropometric, biochemical, haematological and histopathological parameters were also assessed using standard procedures. Results showed that the LD₅₀ values for the acute oral and intraperitoneal toxicity studies for HU were estimated to be 1000 mg/kg and 459.3 mg/kg, respectively. Visible signs of immediate and delayed toxicities including starry hair coat, respiratory distress, and dyskinesia were observed. For the chronic oral toxicity study, HU administered for 90 days produced significant (p<0.001) reductions in the weight gain pattern and significant (p<0.001) and dose related increases in the relative weights of liver, stomach, spleen, testis, lungs and heart, at the 100 and 500 mg/kg of HU. Chronic HU treatment also produced significant (p<0.05, p<0.001) dose related reductions in the serum levels of fasting blood glucose, bicarbonate, urea and creatinine while causing non-significant (p>0.05) alterations in the serum levels of sodium, potassium, alaninine transaminase, aspartate transaminase, alkaline phosphatase, total and conjugated bilirubin, total protein and albumin. Also, chronic oral treatment with HU produced significant (p<0.05, p<0.01, p<0.001) and dose-related increases in the red cell count, packed cell volume, haemoglobin concentration, platelet count, total leucocyte count and lymphocyte differential while producing significant (p<0.05) reductions in neutrophil and granulocyte differentials. HU also produced histological features of proliferations of the stomach epithelia, lung tissues, splenic white and red pulps, and testicular spermatogenic series. Following 14 days of oral toxicity reversibility test, there was no significant (p>0.05) reversal in the serum levels of the biochemical and haematological parameters investigated, including the HU-induced histological lesions.Overall, results of this study showed that HU has a relatively low oral toxicity profile but its prolonged use, particularly, at high doses should be with great caution.Key words: Hunteria umbellata, Toxicity and reversibility profile, Haematology, Liver and Renal function tests, Histopathology, Rodents