Anti-Integrin Therapy To Dampen Inflammation And Reduce Scar Formation

The Role of Inflammation in Scar Formation

Discuss abou the Anti-integrin therapy to dampen Inflammation and reduce scar formation.

Anti- integrin therapy is being widely used by the scientists and the researchers presently. It is   being used in treating several types of diseases such as cancer and help in the survival of human beings. Integrin proteins expressed on both endothelial and tumor cells and  hence it helps in mainly three activities such as  antiangiogenesis; anti-invasion; and anti-tumor.Anti- integrin therapy is also used in curing individuals suffering from including multiple sclerosis, inflammatory bowel disease, and psoriasis. Alpha4beta1 (very late antigen-4) and alphaLbeta2 (leucocyte function-associated antigen-1) integrins have important roles in other immune processes such as including the formation of the immune synapse and the differentiation of T helper 1 lymphocytes.It is very painful for the individuals who suffer from scarring in excess that results from  resulting from burns, scalds and several other injuries.It leads to both physically and psychological effects in them.Presently we do not have any licensed drugs that prevent or  reduce the formation of scar.Inflammation plays a significant role in the normal  of wound healing process.It has been observed that it does not require infection to occur. But if the inflammation is prolonged, it does not lead to the healing of wound (Mosli, Rivera-Nieves and Feagan 2014). Inflammation in excess plays a significant role in the formation of scar in which the level of inflammation in an acute wound will determine the extent to which fibrosis and scarring would occur.

It has been observed through several research experiments that unfavourable  effects  of repair in the tissues  including fibrosis and the formation of scar occurs due inflammation that is  associated with macrophages. According to the experiments, wounds that do not have or have low numbers of macrophages have experienced reduced fibrosis and scarring as compared to wild-type mice.Targeted depletion of macrophages during the early inflammatory stage (days 0-5) of healing resulted in scarring to a minimum level in a mouse model.According to these results it can be suggested that reducing the numbers of macrophages and early inflammation (day 2-5) could potentially reduce the formation of scars.

Macrophages are immune cells  that play a significant role in regulating  the formation of scars. They enter the wound as a monocyte 2 days after the injury occurs.There are a few proteins known as surface integrins  that play an important role for the  migration of  monocytes into sites of inflammation and injury. Integrins are adhesion receptors and  consist of  two subunits, that  help in mediating the cell-cell and cell-matrix interactions. These interactions are important for the   adhesion and migration of the cell. There are different types of integrins and are expressed in a different manner. Integrins such as αDβ2, αMβ2, αXβto, αLβ2 and αEβ7 are specific two leukocytes.

The Role of Macrophages and Integrins in Scar Formation

Integrins  act as excellent therapeutic targets since their ligand-binding domain is exposed to the extracellular milieu and they can be inhibited through pharmacological means.There are certain diseases which can be treated in an effective manner by using licensed anti-integrin therapies that target integrins that are specific to immunity. They are used effectively in clinical settings to treat diseases in which inflammation plays a significant role in the progression of disease.As the inflammatory processes play a crucial role in the healing of wounds and anti-integrin therapies have been successful in reducing inflammation in other diseases, it is proposed that anti-integrin therapy will reduce inflammation and thus improve the formation of scars.

Tumor necrosis factor-α (TNF-α) is a proinflammatory cytokine that is  produced by macrophages, and by renal mesangial and tubular epithelial cells. It  plays a crucial role in stimulating the release of interleukin (IL)-1β, monocyte chemoattractant protein-1 (MCP-1), and transforming growth factor-β (TGF-β). Blockage of TNF-α is presently used in the treatment  of several autoimmune inflammatory diseases after being researched widely in clinical laboratories.  It was hypothesized that blocking TNF-α with a monoclonal antibody would help in preventing inflammation and renal fibrosis in crescentic glomerulonephritis. Thus, the objective of this study is to determine whether blocking specific monocyte integrins reduces inflammation and improves scarring outcomes in a burn mouse model. We have developed a novel therapeutic monoclonal antibody named “Scarless”. In this study we will test the efficiency and mechanisms of Scarless to reduce monocyte migration into burns wound tissue.

Laboratory animals  should be provided with humane care and healthy conditions during their stay in the animal  house of  the laboratory or the institution.The rats used for the experiment should be provided proper food and diet and they should not be harmed or tortured in any manner. All the ethical principles should be followed by the researchers while conducting the experiment and the members of the ethical committee should keep a check on the procedures that are being followed. The facilities of  housing the animals in the laboratory should meet or exceed current regulations and guidelines and should be inspected twice a year.There should be experienced psychologists , veterinary doctors  and their assistants should  keep a check on the behavior  of the animals who are taken into consideration for the experiment. They should keep a check and be aware of the differences between unusual behaviors and the normal.They should assess them in a proper manner and provide treatment of health problems if required. 

Anti-Integrin Therapy in Reducing Inflammation and Scarring

Two groups of rats are taken for the study and one of them acts as the control to compare the results at the end of the experiment.

Materials of Immuno histochemistry such as slides, paraffin sections and stains

Antibody blockade of TNF-a reduces inflammation and scarring .Male rats of weight between 200 to 250 grams  and age between 6-8 weeks  should be taken .They should be provided proper diet and water (Schulz et al.2015)A blocking IgG2a monoclonal antibody to rat TNFa (C432) should be used in the experiment. Immunohistochemistry and Immunofluorosence technique is to be used in the procedure. The monoclonal antibodies  that are  specific for rat monocytes or  macrophages should used while conducting the experiment. Nephrotoxic nephritis was induced in rats by intravenous injection of rabbit antirat glomerular basement membrane (GBM) nephrotoxic serum (NTS). Anti-TNF-α monoclonal antibody or saline was given  by intraperitoneal means and three times per week in four protocols: experiment 1, days 0 to 7; experiment 2, days 0 to 14 and days 4 to 14; experiment 3, days 4 to 28; and experiment 4, days 14 to 28.

In the experiment 1, rats that were  treated  from disease induction had less glomerular fibrinoid necrosis and fewer glomerular macrophages at day 7. In the  experiment 2, rats treated from day 0 or day 4 showed an improvement in the  renal function, as  observed by serum creatinine, with a significant reduction in crescents.In experiment 3, anti-TNF-α treatment reduced urine protein to creatinine ratio and urinary MCP-1 levels by a significant amount. Serum creatinine was preserved at both the days 14 and day 28. Tubulointerstitial inflammation, glomerular and tubulointerstitial scarring, and markers of fibrosis [α-smooth muscle actin (α-SMA) and type IV collagen] were less  in treated rats at day 28 in a significant manner. In the experiment 4, serum creatinine was  greater and tubulointerstitial scarring was lower in delayed-treated animals.

Conclusion

When the endogenous TNF-α is neutralized it leads to a reduction in inflammation in the glomerulus, formation of crescent, and tubulointerstitial scarring, with preservation of renal function, in experimental crescentic glomerulonephritis. Blockade of TNF-α is effective even when it is introduced at the time of maximum glomerular inflammation.After the study, it can be concluded that blocking TNF-a prevents inflammatory response in mouse model (Berghe et al.2014 ).

References

Berghe, T.V., Linkermann, A., Jouan-Lanhouet, S., Walczak, H. and Vandenabeele, P., 2014. Regulated necrosis: the expanding network of non-apoptotic cell death pathways. Nature reviews. Molecular cell biology, 15(2), p.135.

Freeman, S.A., Goyette, J., Furuya, W., Woods, E.C., Bertozzi, C.R., Bergmeier, W., Hinz, B., Van Der Merwe, P.A., Das, R. and Grinstein, S., 2016. Integrins form an expanding diffusional barrier that coordinates phagocytosis. Cell, 164(1), pp.128-140.

Ley, K., Rivera-Nieves, J., Sandborn, W.J. and Shattil, S., 2016. Integrin-based therapeutics: biological basis, clinical use and new drugs. Nature reviews. Drug discovery, 15(3), p.173.

Mosli, M.H., Rivera-Nieves, J. and Feagan, B.G., 2014. T-cell trafficking and anti-adhesion strategies in inflammatory bowel disease: current and future prospects. Drugs, 74(3), pp.297-311.

Schulz, J.N., Zeltz, C., Sørensen, I.W., Barczyk, M., Carracedo, S., Hallinger, R., Niehoff, A., Eckes, B. and Gullberg, D., 2015. Reduced granulation tissue and wound strength in the absence of α11β1 integrin.Journal of Investigative Dermatology, 135(5), pp.1435-1444.

Wynn, T.A., Chawla, A. and Pollard, J.W., 2013. Origins and Hallmarks of macrophages: development, homeostasis, and disease. Nature, 496(7446), p.445.