Int J Stem Cells.  2014 Nov;7(2):87-97. 10.15283/ijsc.2014.7.2.87.

Role of Bone Marrow Mesenchymal Stem Cells in the Treatment of CCL4 Induced Liver Fibrosis in Albino Rats: A Histological and Immunohistochemical Study

• Ahmed SK
• Mohammed S
• Khalaf G
• Fikry H

• KMID: 1974596
• DOI: http://doi.org/10.15283/ijsc.2014.7.2.87

Abstract

BACKGROUND AND OBJECTIVES
Variety of pathological factors including viral hepatitis, alcohol and drug abuse, metabolic diseases, autoimmune diseases and congenital abnormalities can cause hepatic injury. Liver transplantation is the treatment of choice for end-stage liver diseases, however, it faces several difficulties. So the aim of the work is to evaluate the effect of bone marrow derived mesenchymal stem cells (BM-MSCs) on the liver structure in carbon tetra chloride CCL4 induced liver fibrosis in rats. MATERIALS AND RESULTS: BM-MSCs were isolated and characterized from long bones of twenty male albino rats. Sixty female rats were divided into the following two groups: Group I; thirty rats which were the control group. Group II; thirty rats were injected intra-peritoneal (IP) by CCL4 twice weekly for four weeks and was further subdivided into the following three subgroups: Subgroup IIA (CCL4 alone); included ten rats which were sacrificed after this four weeks. Subgroup IIB (CCL4/MSCs); included ten rats which were IP injected by a single dose of BM-MSCs and were sacrificed after four weeks. Subgroup IIC (CCL4/recovery); included ten rats which were left for another four weeks without any intervention. Histological examination of liver specimens showed that CCl4 caused variable pathological changes with elevated liver enzymes. Injection of BM-MSCs revealed an improvement in the histological picture of the liver and its enzymatic profile. On the other hand, most of the pathological lesion were still detected in rats of recovery group.
CONCLUSIONS
BM-MSC could restore the liver structure and function in experimental model of liver fibrosis.

Keyword

Bone Marrow; Liver; Fibrosis; CCL4; Histopathology; MSCs

MeSH Terms

Animals
Autoimmune Diseases
Bone Marrow*
Carbon
Characidae
Congenital Abnormalities
Female
Fibrosis
Hand
Hepatitis
Humans
Liver
Liver Cirrhosis*
Liver Diseases
Liver Transplantation
Male
Mesenchymal Stromal Cells*
Metabolic Diseases
Models, Theoretical
Rats*
Substance-Related Disorders
Carbon

Figure

• 

  Fig. 1. (A) Showing radiating cords of hepatocytes from the central vein (C.V.). The hepatocytes have central, rounded, vesicular nuclei (↑) and acidophilic cytoplasm. Some of the cells appear bi-nucleated (▲). Notice the lining cells ( ) of the blood sinusoids (control H and E. ×720). (B) Most of the hepatocytes are vacuolated ( ). Few hepatocytes appear with acidophilic cytoplasm and deeply stained nuclei (▲) (CCL4 alone group. H and E. ×560). (C) Most of the hepatocytes have granular acidophilic cytoplasm and vesicular nuclei. Few cells show cytoplasmic vacuolation (↑) (CCL4/MSCs group. H and E. ×560). (D) Showing highly vacuolated hepatocytes with deeply stained nuclei (↑) (CCL4/Recovery group, H and E. ×400).

• 

  Fig. 2. (A) Showing few collagen fibers ( ) surrounding the central vein (V), in portal area (P) and in the capsule (C) in Control group. (B) Showing numerous collagen fibers ( ) surrounding the central veins (V), in portal area (P) and in the capsule (C) in CCL4 alone group. (C) Showing the collagen fibers of CCL4/MSCs group which are comparable to that of the control group. (D) Showing numerous collagen fibers (↑) surrounding the central veins (V), in portal area (P) and in the capsule (C) in CCL4/Recovery group (Masson’s trichrome V×400, P×400, C×140).

• 

  Fig. 3. (A) Showing few α-SMA positive cells ( ) around the central vein and in-between the hepatocytes (control group×560). (B) Showing an apparent increase in α-SMA positive cells ( ) around central vein, in septa between the hepatic lobules and in-between the hepatocytes. Inset: α-SMA positive cells appear spindle in shape with cytoplasmic processes (CCL4 alone group×400–inset×560). (C) Showing few α-SMA positive cells ( ) around the central vein and in-between the hepatocytes (CCL4/MSCs group×560). (D) Showing strong positive immune reaction for α-SMA ( ) around the central vein and in-between the hepatocytes (CCL4/Recovery group×400). Immunostaining for α-SMA.

• 

  Fig. 4. (A) Showing few hepatocytes with positive immune reaction for PCNA ( ) in control group. (B) Showing few hepatocytes with positive immune reaction for PCNA ( ) in CCL4 alone group. (C) Showing many of PCNA positive hepatocytes ( ) in CCL4/MSCs group. (D) Showing few hepatocytes with positive immune reaction for PCNA ( ) in CCL4/Recovery group (Immunostaining for PCNA, ×560).

• 

  Fig. 5. (A) Showing adjacent hepatocytes with sinusoidal spaces (S) in-between. HSCs (*) are seen in the perisinusoidal space The hepatocytes appear with large rounded, central vesicular nuclei (N). The nuclei show usual characteristic chromatin distribution. The cytoplasm contains numerous mitochondria (M), rER ( ) and fat droplets (▲) (Control group TEM×4050). (B) Showing adjacent hepatocytes. The nuclei of the hepatocytes show abnormal chromatin distribution (N). The cytoplasm contains multiple vacuoles ( ) and fat droplets (▲) (CCL4 alone group TEM ×4050).

• 

  Fig. 6. (A) Showing hepatocytes (white ) that are comparable to that of the control group. They contain many mitochondria and show usual distribution of chromatin in their nuclei (N). Small vacuoles ( ) and few fat droplets (▲) are seen in some hepatocytes (CCL4/MSCs group TEM×4050). (B) Showing irregular large vacuoles (▲) and fat droplets ( ) inside the cytoplasm of hepatocytes. The nuclei appear with abnormal chromatin distribution (N). Notice the presence of HSCs (*) with euchromatic nuclei in between hepatocytes (CCL4/Recovery group TEM×4050).

• 

  Fig. 7. Showing presence of Y chromosomes in male rats from which BM-MSCs were isolated (lane 1) and female rats treated with BM-MSCs in CCL4/MSCs group (lane 4). Y chromosome marker is not detecting in female rats of the control group (lane 2), CCL4 alone group (lane 3) and CCL4/recovery group (lane 5). M: PCR marker (U.V. trans-illuminated agarose gel of PCR products of SRY gene).

Reference

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