Tissue regeneration and stem cell distribution in adriamycin induced glomerulopathy

Abstract

Background and objectives: Glomerulosclerosis develops secondary to various kidney diseases. It was postulated that adriamycin (ADR) induce chronic glomerulopathy. Treatment combinations for one year did not significantly modify renal function in resistant focal segmental glomerulosclerosis (FSGS). Recurrence of FSGS after renal transplantation impacts long-term graft survival and limits access to transplantation. The present study aimed at investigating the relation between the possible therapeutic effect of human mesenchymal stem cells (HMSCs), isolated from cord blood on glomerular damage and their distribution by using ADR induced nephrotoxicity as a model in albino rat.

Methods and results: Thirty three male albino rats were divided into control group, ADR group where rats were given single intraperitoneal (IP) injection of 5 mg/kg adriamycin. The rats were sacrificed 10, 20 and 30 days following confirmation of glomerular injury. In stem cell therapy group, rats were injected with HMSCs following confirmation of renal injury and sacrificed 10, 20 and 30 days after HMSCs therapy. Kidney sections were exposed to histological, histochemical, immunohistochemical, morphometric and serological studies. In response to SC therapy multiple Malpighian corpuscles (MC) appeared with patent Bowman's space (Bs) 10 and 20 days following therapy. One month following therapy no remarkable shrunken glomeruli were evident. Glomerular area and serum creatinine were significantly different in ADR group in comparison to control and SC therapy groups.

Conclusions: ADR induced glomerulosclerosis regressed in response to cord blood HMSC therapy. A reciprocal relation was recorded between the extent of renal regeneration and the distribution of undifferentiated mesenchymal stem cells.

Keywords: Adriamycin; Cord blood; Glomerulosclerosis; Mesenchymal stem cells.

Fig. 1.. Malpighian renal corpuscle containing a glomerulus (g), Bowman's space (Bs), parietal layer of Bowman's capsule (arrow), proximal convoluted tubules (PCT) (p) and distal convoluted tubules (DCT) (d) in control group (renal cortex; H&E, ×200).

Fig. 2.. A corpuscle with completely obliterated Bs (thick arrow), a second with partially obliterated space (arrowhead) and a third with patent space (*). Note vacuolated cytoplasm and dark nuclei (thin arrows) of the cells lining multiple cortical tubules in subgroup A1 (renal cortex; H&E, ×200).

Fig. 3.. A corpuscle with completely obliterated Bowman' space (thick arrow) and two corpuscles with partially obliterated spaces (arrowheads). Note vacuolated cytoplasm and dark nuclei (thin arrows) of the cells lining multiple cortical tubules in subgroup A2 (renal cortex; H&E, ×200).

Fig. 4.. Two corpuscles with shrunken glomeruli and wide Bowman' space (*). Note a corpuscle with separated glomerular loops (curved arrow), empty space lined by squamous cells (thick arrow), vacuolated cytoplasm of the cells lining multiple cortical tubules (thin arrows) and detached epithelial lining (arrowhead) of a cortical tubule in subgroup A3 (renal cortex; H&E, ×200).

Fig. 5.. A corpuscle with patent space (*). Vacuolated cytoplasm and dark nuclei (thin arrows) of the cells lining few cortical tubules are seen. Note mononuclear infiltration (arrowhead) in subgroup S1 (renal cortex; H&E, ×200).

Fig. 6.. Five corpuscles with patent space (*). Note vacuolated cytoplasm and dark nuclei (thin arrows) of the cells lining few cortical tubules in subgroup S2 (renal cortex; H&E, ×200).

Fig. 7.. Six corpuscles with patent Bowman's space (*) and few tubules with vacuolated cytoplasm and dark nuclei of the lining cells (thin arrows) in subgroup S3. Sections in the renal cortex showing (renal cortex; H&E, ×200).

Fig. 8.. Multiple spindle (s) and few cuboidal (cu) Pb+ve cells at the glomerulus, in the Bs and at the epithelial lining of the cortical tubules in subgroup S1 (renal cortex; Prussian blue, ×400).

Fig. 9.. Multiple spindle (s) and few cuboidal (cu) Pb+ve cells in peritubular capillaries and among the epithelial lining of CT in subgroup S1 (renal cortex; Prussian blue, ×400).

Fig. 10.. Two cuboidal (cu) and fewer spindle (s) +ve cells at the glomerulus, in the Bs and at the lining epithelial cells of the cortical tubules in subgroup S3 (renal cortex; Prussian blue, ×400).

Fig. 11.. +ve immunostaining in multiple cortical tubules (arrows) in subgroup S1 (CD105, ×200).

Fig. 12.. Higher magnification of the previous figure showing granular reaction in multiple spindle cells (arrows) among the lining of cortical tubules (CD105, ×1,000).

Fig. 13.. +ve immunostaining in fewer cortical tubules (arrows) in subgroup S2 compared to Fig. 11 (CD105, ×200).

Fig. 14.. +ve immunostaining in fewer cortical tubules (arrows) compared to Figs. 11 and 13 in subgroup S3 (CD105, ×200).

Fig. 15.. Mean glomerular area in control and experimental groups.

Fig. 16.. Mean area% of Prussian blue positive cells in control and experimental groups.

Fig. 17.. Mean area% of CD105 positive cells in control and experimental groups.

Fig. 18.. Mean value of serum creatinine in control and experimental groups.