Isolation and Assessment of Mesenchymal Stem Cells Derived From Bone Marrow: Histologic and Histomorphometric Study in a Canine Periodontal Defect

Abstract

The aim of the present study was to investigate an isolation procedure to culture mesenchymal stem cells derived from bone marrow and evaluate their potential in periodontal regeneration. Potential stem cells from bone marrow, aspirated from the iliac crest of nine mongrel canines 1 to 2 years of age, were cultivated. After the examination of surface epitopes of the isolated cells, the total RNA from osteogenic, adipogenic, and chondrogenic cell cultures were analyzed by reverse transcription polymerase chain reaction (RT-PCR) to confirm stem cell gene expressions. 2 × 10(7) mL of the stem cells were loaded on 0.2 mL of anorganic bovine bone mineral (ABBM) granules. In each animal, bilateral acute/chronic intrabony periodontal defects were created surgically and by placement of ligatures around the cervical aspect of the teeth. At week 5, after flap debridement, the bilateral defects were randomly assigned to 2 treatment groups: the control group received ABBM, and the test group received BMSCs-loaded ABBM. Eight weeks after transplantation, regenerative parameters were analyzed histologically and histometrically. The RNA expressions confirmed the cultivation of mesenchymal stem cell. More new cementum and periodontal ligament (PDL) were measured in the test group (cementum: 3.33 ± 0.94 vs 2.03 ± 1.30, P = 0.027; PDL: 2.69 ± 0.73 vs 1.53 ± 1.21, P = 0.026). New bone formation was similar in both groups (2.70 ± 0.86 vs 1.99 ± 1.31; P = 0.193). Mesenchymal stem cells derived from bone marrow should be considered a promising technique for use in patients with periodontal attachment loss and merits further investigations.

Efficacy of stem cells on periodontal regeneration: Systematic review of pre‐clinical studies

“…Ten of 22 studies reported quantitative data on histological evidence of new bone, cementum and PDL in the periodontal defect . Most of the remaining studies reported histological quantitative data restricted to new bone and cementum .…”

“…Most studies (14 of 22) used dogs and six studies used rats . Finally, merino sheep and minipigs were used in only one study each.…”

“…Periodontal defects were surgically created in all studies. Differences in defect morphology were identified among the studies that evaluated: three‐wall intrabony defects, one‐wall intrabony defects, class III and class II furcations, fenestrations, dehiscences, circumferential and supra‐alveolar defects. In some of these studies, following surgical defect creation, defects were filled with anaerobic bacteria or impression materials to induce chronic inflammation.…”

“…In some of these studies, following surgical defect creation, defects were filled with anaerobic bacteria or impression materials to induce chronic inflammation. In others, ligatures were secured at the cervical aspect of affected teeth to allow plaque accumulation after surgical defect creation …”

“…Autogenous, allogenous and xenogenous MSCs were used to treat periodontal defects. These encompassed nine different types/sources of MSC.…”