Effect of Bone Marrow Stem Cells on the Healing of Colonic Anastomosis in Rats

Veracious, Cornerstone Wann (2008) Effect of Bone Marrow Stem Cells on the Healing of Colonic Anastomosis in Rats. Masters thesis, Christian Medical College, Vellore.


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INTRODUCTION: The concept of cell regeneration dates well back to ancient history, when Prometheus transgressed the Laws of the ancient gods and stole fire for mankind. He was chained to Mount Caucasus, where a vulture preyed on his liver daily which renewed (regenerate) as quickly as it was devoured1. Wound healing is a never ending process in the human body. Cells in the skin, lungs, intestinal mucosa regenerate everyday from self renewing progenitors cell located at specific locations in that organ (eg, intestinal mucosal crypts). These nests of progenitor cells are called niches (Fr.recess). The progenitor cells in tissue organs are mesenchymal stem cells. The concept of organ specific regeneration where cells give rise to their own ‘type’ has been challenged. These mesenchymal stem cells can be multipotent, resembling pluripotent cell population derived from embryos. Plasticity, the ability of a stem cell of a particular cell line to differentiate into a cell of different germinal layer (eg, bone marrow, a mesoderm into nerve tissue, an ectoderm and vice versa) has enabled researchers to explore this potential of tissue repair in spinal cord injury and acute myocardial infarction. Colonic anastamosis is a commonly performed operation in general surgery. Despite improvements in suture materials, techniques, mechanical aid and various nutritional factors, intestinal anastamosis continues to be complicated by dehiscence and stricture formation. Anastamotic dehiscence following colorectal operation is a significant cause of morbidity and mortality in the perioperative period. Low colonic anastomoses have clinically significant leakage rates of around 16% with radiologically demonstrable ones ranging from 27 – 49% Various factors involved in the healing process are not completely understood3. This study proposes to look at the role of bone marrow cells in the healing of colonic anastamosis in rats. Mesenchymal stem cells derived from bone marrow has been used extensively in bone marrow transplants for various indications. Cultured mesenchymal cells ensure the role of stem cells in the healing process but bone marrow cells can be directly innoculated into injured tissue without culture as marrow tissue has a high concentration of multipotent hemopoietic cells. AIMS AND OBJECTIVES: To assess whether Bone marrow derived stem cells when injected into colonic anastomosis: 1. Provide adequate cells and response for repair. 2. Improve the mechanical strength of colonic anastomosis. MATERIAL AND METHODS: A clearance from the institutional animal ethics committee was obtained prior to conducting the experimental study. 20 female and 5 male albino rats of the Wistar strain weighing between 150 to 250 gms were used for the experiment. Bone marrow was harvested from the long bones of one male rat and preserved in a media (GIBCO™ Iscove's Modified Dulbecco's Medium (IMDM) (1X) liquid containing L-glutamine, 25 mM HEPES buffer, 3,024 mg/L sodium bicarbonate) for the experiment on 5 to 6 female rats. The experiment was conducted on female rats so that the present of the donor stem cells could be detected later by identifying Y- chromosomes at the site of anastomosis using Fluorescence in situ hybridization (FISH) probes. Randomization: Two colonic anastamoses were done on each animal, one proximal and the other distal. The prepared bone marrow suspension was injected into one of the anastomosis and a media alone was injected into the other according to the randomization table .The rats were then sacrificed. Bursting pressure was done on alternate rats and histological examination done on the others. The pathologist was blinded. CONCLUSIONS: 1. Stem cells derived from the bone marrow when injected at the site of anastomosis did not significantly alter the bursting pressure or histology at the site of anastamosis. 2. There was more inflammation when bone marrow derived stem cells were injected thereby indicating an increased risk of leak. 3. There was less fibrosis in the bone marrow derived stem cell injected group of anastamosis as compared to the control group. 10.0 Clinical Significance and Role for Further Research This is the first study done using crude stem cells derived from the bone marrow. A study done with cultured stem cells may be more appropriate. Technical perfection in harvesting and culturing stem cells can be achieved and this may help in future studies. The use of Fluorescence in situ hybridization (FISH) probe in identification of the donor cells would help in consolidating the effectiveness of the use stem cells in this study and or similar studies.

Item Type: Thesis (Masters)
Uncontrolled Keywords: Bone Marrow Stem Cells ; Healing ; Colonic Anastomosis ; Rats.
Subjects: MEDICAL > General Surgery
Depositing User: Devi S
Date Deposited: 04 May 2018 03:28
Last Modified: 04 May 2018 03:28
URI: http://repository-tnmgrmu.ac.in/id/eprint/7426

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