Neural-like cells from adipose-derived stem cells for cavernous nerve injury in rats
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摘要
Although the remaining nerve tissue can regenerate and partly restore erectile function when the cavernous nerve is compressed/severed and function lost, the limited regenerative ability of these nerve tissues often fails to meet clinical needs. Adipose-derived stem cells are easy to obtain and culture, and can differentiate into neural cells. Their proliferation rate is easy to control and they may be used to help restore injured cavernous nerve function. Sprague-Dawley male rats(n = 45) were equally randomized into three groups: fifteen rats as a sham-operated group, fifteen rats as a bilateral nerve crush(BINC) group(with no further intervention), fifteen rats as a BINC with intracavernous injection of one million neural-like cells from adipose-derived stem cells(NAS)(BINC + NAS) group. After 4 weeks, erectile function was assessed by stimulating the cavernous body. The number of myelinated axons in the dorsal cavernous nerve was determined by toluidine blue staining. The area of neuronal nitric oxide synthase-positive fibers in the dorsal penile nerve was measured by immunohistochemical staining. Masson staining was used to analyze the ratio of smooth muscle to collagen in penile tissue. The results demonstrate that maximal intracavernous pressure, the ratio of maximal intracavernous pressure to mean arterial pressure, the numbers of myelinated axons and neuronal nitric oxide synthase-positive fibers in the dorsal penile nerve, and the ratio of smooth muscle to collagen could be increased after cell transplantation. These findings indicate that neural-like cells from adipose-derived stem cells can effectively alleviate cavernous nerve injury and improve erectile function. All animal experiments were approved by the Animal Ethics Committee of Huazhong University of Science and Technology, China(approval No. 2017-1925) on September 15, 2017.
Although the remaining nerve tissue can regenerate and partly restore erectile function when the cavernous nerve is compressed/severed and function lost, the limited regenerative ability of these nerve tissues often fails to meet clinical needs. Adipose-derived stem cells are easy to obtain and culture, and can differentiate into neural cells. Their proliferation rate is easy to control and they may be used to help restore injured cavernous nerve function. Sprague-Dawley male rats(n = 45) were equally randomized into three groups: fifteen rats as a sham-operated group, fifteen rats as a bilateral nerve crush(BINC) group(with no further intervention), fifteen rats as a BINC with intracavernous injection of one million neural-like cells from adipose-derived stem cells(NAS)(BINC + NAS) group. After 4 weeks, erectile function was assessed by stimulating the cavernous body. The number of myelinated axons in the dorsal cavernous nerve was determined by toluidine blue staining. The area of neuronal nitric oxide synthase-positive fibers in the dorsal penile nerve was measured by immunohistochemical staining. Masson staining was used to analyze the ratio of smooth muscle to collagen in penile tissue. The results demonstrate that maximal intracavernous pressure, the ratio of maximal intracavernous pressure to mean arterial pressure, the numbers of myelinated axons and neuronal nitric oxide synthase-positive fibers in the dorsal penile nerve, and the ratio of smooth muscle to collagen could be increased after cell transplantation. These findings indicate that neural-like cells from adipose-derived stem cells can effectively alleviate cavernous nerve injury and improve erectile function. All animal experiments were approved by the Animal Ethics Committee of Huazhong University of Science and Technology, China(approval No. 2017-1925) on September 15, 2017.
Although the remaining nerve tissue can regenerate and partly restore erectile function when the cavernous nerve is compressed/severed and function lost, the limited regenerative ability of these nerve tissues often fails to meet clinical needs. Adipose-derived stem cells are easy to obtain and culture, and can differentiate into neural cells. Their proliferation rate is easy to control and they may be used to help restore injured cavernous nerve function. Sprague-Dawley male rats(n = 45) were equally randomized into three groups: fifteen rats as a sham-operated group, fifteen rats as a bilateral nerve crush(BINC) group(with no further intervention), fifteen rats as a BINC with intracavernous injection of one million neural-like cells from adipose-derived stem cells(NAS)(BINC + NAS) group. After 4 weeks, erectile function was assessed by stimulating the cavernous body. The number of myelinated axons in the dorsal cavernous nerve was determined by toluidine blue staining. The area of neuronal nitric oxide synthase-positive fibers in the dorsal penile nerve was measured by immunohistochemical staining. Masson staining was used to analyze the ratio of smooth muscle to collagen in penile tissue. The results demonstrate that maximal intracavernous pressure, the ratio of maximal intracavernous pressure to mean arterial pressure, the numbers of myelinated axons and neuronal nitric oxide synthase-positive fibers in the dorsal penile nerve, and the ratio of smooth muscle to collagen could be increased after cell transplantation. These findings indicate that neural-like cells from adipose-derived stem cells can effectively alleviate cavernous nerve injury and improve erectile function. All animal experiments were approved by the Animal Ethics Committee of Huazhong University of Science and Technology, China(approval No. 2017-1925) on September 15, 2017.
引文
Albersen M,Fandel TM,Lin G,Wang G,Banie L,Lin CS,Lue TF(2010)Injections of adipose tissue-derived stem cells and stem cell lysate improve recovery of erectile function in a rat model of cavernous nerve injury.J Sex Med 7:3331-3340.
An C,Cheng Y,Yuan Q,Li J(2010)IGF-1 and BMP-2 induces differentiation of adipose-derived mesenchymal stem cells into chondrocytes-like cells.Ann Biomed Eng 38:1647-1654.
Bochinski D,Lin GT,Nunes L,Carrion R,Rahman N,Lin CS,Lue TF(2004)The effect of neural embryonic stem cell therapy in a rat model of cavernosal nerve injury.BJU Int 94:904-909.
Burnett AL,Musicki B(2005)The nitric oxide signaling pathway in the penis.Curr Pharm Des 11:3987-3994.
Carrier S,Zvara P,Nunes L,Kour NW,Rehman J,Lue TF(1995)Regeneration of nitric oxide synthase-containing nerves after cavernous nerve neurotomy in the rat.J Urol 153:1722-1727.
Chen F,Zhang H,Wang Z,Ding W,Zeng Q,Liu W,Huang C,He S,Wei A(2017)Adipose-derived stem cell-derived exosomes ameliorate erectile dysfunction in a rat model of type 2 diabetes.J Sex Med 14:1084-1094.
Chen J,Tang YX,Liu YM,Chen J,Hu XQ,Liu N,Wang SX,Zhang Y,Zeng WG,Ni HJ,Zhao B,Chen YF,Tang ZP(2012)Transplantation of adipose-derived stem cells is associated with neural differentiation and functional improvement in a rat model of intracerebral hemorrhage.CNS Neurosci Ther 18:847-854.
Dubbelman YD,Dohle GR,Schr?der FH(2006)Sexual function before and after radical retropubic prostatectomy:A systematic review of prognostic indicators for a successful outcome.Eur Urol 50:711-720.
Fandel TM,Albersen M,Lin G,Qiu X,Ning H,Banie L,Lue TF,Lin CS(2012)Recruitment of intracavernously injected adipose-derived stem cells to the major pelvic ganglion improves erectile function in a rat model of cavernous nerve injury.Eur Urol 61:201-210.
Fansa H,Keilhoff G(2003)Factors influencing nerve regeneration.Handchir Mikrochir Plast Chir 35:72-82.
Fernandes M,Valente SG,Sabongi RG,Gomes Dos Santos JB,Leite VM,Ulrich H,Nery AA,da Silva Fernandes MJ(2018)Bone marrow-derived mesenchymal stem cells versus adipose-derived mesenchymal stem cells for peripheral nerve regeneration.Neural Regen Res 13:100-104.
Heidenreich A,Bellmunt J,Bolla M,Joniau S,Mason M,Matveev V,Mottet N,Schmid HP,van der Kwast T,Wiegel T,Zattoni F,European Association of Urology(2011)EAU guidelines on prostate cancer.Part 1:screening,diagnosis,and treatment of clinically localised disease.Eur Urol 59:61-71.
Jang S,Cho HH,Cho YB,Park JS,Jeong HS(2010)Functional neural differentiation of human adipose tissue-derived stem cells using bFGF and forskolin.BMC Cell Biol 11:25.
Kato H,Minamizato H,Ohno H,Ohira Y,Izawa T(2019)Exercise ameliorates high-fat diet-induced impairment of differentiation of adipose-derived stem cells into neuron-like cells in rats.J Cell Physiol 234:1452-1460.
Lin G,Banie L,Ning H,Bella AJ,Lin CS,Lue TF(2009)Potential of adipose-derived stem cells for treatment of erectile dysfunction.J Sex Med 6Suppl 3:320-327.
Lin G,Albersen M,Harraz AM,Fandel TM,Garcia M,McGrath MH,Konety BR,Lue TF,Lin CS(2011)Cavernous nerve repair with allogenic adipose matrix and autologous adipose-derived stem cells.Urology77:1509.e1-8.
Lu P,Jones LL,Snyder EY,Tuszynski MH(2003)Neural stem cells constitutively secrete neurotrophic factors and promote extensive host axonal growth after spinal cord injury.Exp Neurol 181:115-129.
Madonna R,Geng YJ,De Caterina R(2009)Adipose tissue-derived stem cells:characterization and potential for cardiovascular repair.Arterioscler Thromb Vasc Biol 29:1723-1729.
Makdissy N,Haddad K,AlBacha JD,Chaker D,Ismail B,Azar A,Oreibi G,Ayoub D,Achkar I,Quilliot D,Fajloun Z(2018)Essential role of AT-P6AP2 enrichment in caveolae/lipid raft microdomains for the induction of neuronal differentiation of stem cells.Stem Cell Res Ther 9:132.
Nagasaki H,Shang Q,Suzuki T,Hashimoto H,Yoshimura T,Kondo TA,Ozaki T,Maruyama S,Jomori T,Oiso Y,Hamada Y(2011)Low-serum culture system improves the adipogenic ability of visceral adipose tissue-derived stromal cells.Cell Biol Int 35:559-568.
Patrick CW,Jr.(2000)Adipose tissue engineering:the future of breast and soft tissue reconstruction following tumor resection.Semin Surg Oncol19:302-311.
Qian DX,Zhang HT,Ma X,Jiang XD,Xu RX(2010)Comparison of the efficiencies of three neural induction protocols in human adipose stromal cells.Neurochem Res 35:572-579.
Safford KM,Safford SD,Gimble JM,Shetty AK,Rice HE(2004)Characterization of neuronal/glial differentiation of murine adipose-derived adult stromal cells.Exp Neurol 187:319-328.
Safford KM,Hicok KC,Safford SD,Halvorsen YD,Wilkison WO,Gimble JM,Rice HE(2002)Neurogenic differentiation of murine and human adipose-derived stromal cells.Biochem Biophys Res Commun 294:371-379.
Santiago LY,Clavijo-Alvarez J,Brayfield C,Rubin JP,Marra KG(2009)Delivery of adipose-derived precursor cells for peripheral nerve repair.Cell Transplant 18:145-158.
Sun Y,Jin K,Xie L,Childs J,Mao XO,Logvinova A,Greenberg DA(2003)VEGF-induced neuroprotection,neurogenesis,and angiogenesis after focal cerebral ischemia.J Clin Invest 111:1843-1851.
Walsh PC(1987)Radical prostatectomy,preservation of sexual function,cancer control.The controversy.Urol Clin North Am 14:663-673.
Walz J,Burnett AL,Costello AJ,Eastham JA,Graefen M,Guillonneau B,Menon M,Montorsi F,Myers RP,Rocco B,Villers A(2010)A critical analysis of the current knowledge of surgical anatomy related to optimization of cancer control and preservation of continence and erection in candidates for radical prostatectomy.Eur Urol 57:179-192.
Ying C,Hu W,Cheng B,Zheng X,Li S(2012)Neural differentiation of rat adipose-derived stem cells in vitro.Cell Mol Neurobiol 32:1255-1263.
Ying C,Yang M,Zheng X,Hu W,Wang X(2013)Effects of intracavernous injection of adipose-derived stem cells on cavernous nerve regeneration in a rat model.Cell Mol Neurobiol 33:233-240.
Zarei MH,Soodi M,Qasemian-Lemraski M,Jafarzadeh E,Taha MF(2016)Study of the chlorpyrifos neurotoxicity using neural differentiation of adipose tissue-derived stem cells.Environ Toxicol 31:1510-1519.
Zhang H,Yang R,Wang Z,Lin G,Lue TF,Lin CS(2011)Adipose tissue-derived stem cells secrete CXCL5 cytokine with neurotrophic effects on cavernous nerve regeneration.J Sex Med 8:437-446.
Zhang HT,Luo J,Sui LS,Ma X,Yan ZJ,Lin JH,Wang YS,Chen YZ,Jiang XD,Xu RX(2009)Effects of differentiated versus undifferentiated adipose tissue-derived stromal cell grafts on functional recovery after spinal cord contusion.Cell Mol Neurobiol 29:1283-1292.
Zhao ZK,Yu HL,Liu B,Wang H,Luo Q,Ding XG(2016)Antioxidative mechanism of Lycium barbarum polysaccharides promotes repair and regeneration following cavernous nerve injury.Neural Regen Res11:1312-1321.
Zheng Y,Huang C,Liu F,Lin H,Yang X,Zhang Z(2017)Comparison of the neuronal differentiation abilities of bone marrow derived and adipose tissuederived mesenchymal stem cells.Mol Med Rep 16:3877-3886.
Zhong B,Liu X,Wu W(2018)Transplantation of neural stem cells from different sources in the treatment of cerebral ischemia injury in rats.Zhongguo Zuzhi Gongcheng Yanjiu 22:3349-3356.
An C,Cheng Y,Yuan Q,Li J(2010)IGF-1 and BMP-2 induces differentiation of adipose-derived mesenchymal stem cells into chondrocytes-like cells.Ann Biomed Eng 38:1647-1654.
Bochinski D,Lin GT,Nunes L,Carrion R,Rahman N,Lin CS,Lue TF(2004)The effect of neural embryonic stem cell therapy in a rat model of cavernosal nerve injury.BJU Int 94:904-909.
Burnett AL,Musicki B(2005)The nitric oxide signaling pathway in the penis.Curr Pharm Des 11:3987-3994.
Carrier S,Zvara P,Nunes L,Kour NW,Rehman J,Lue TF(1995)Regeneration of nitric oxide synthase-containing nerves after cavernous nerve neurotomy in the rat.J Urol 153:1722-1727.
Chen F,Zhang H,Wang Z,Ding W,Zeng Q,Liu W,Huang C,He S,Wei A(2017)Adipose-derived stem cell-derived exosomes ameliorate erectile dysfunction in a rat model of type 2 diabetes.J Sex Med 14:1084-1094.
Chen J,Tang YX,Liu YM,Chen J,Hu XQ,Liu N,Wang SX,Zhang Y,Zeng WG,Ni HJ,Zhao B,Chen YF,Tang ZP(2012)Transplantation of adipose-derived stem cells is associated with neural differentiation and functional improvement in a rat model of intracerebral hemorrhage.CNS Neurosci Ther 18:847-854.
Dubbelman YD,Dohle GR,Schr?der FH(2006)Sexual function before and after radical retropubic prostatectomy:A systematic review of prognostic indicators for a successful outcome.Eur Urol 50:711-720.
Fandel TM,Albersen M,Lin G,Qiu X,Ning H,Banie L,Lue TF,Lin CS(2012)Recruitment of intracavernously injected adipose-derived stem cells to the major pelvic ganglion improves erectile function in a rat model of cavernous nerve injury.Eur Urol 61:201-210.
Fansa H,Keilhoff G(2003)Factors influencing nerve regeneration.Handchir Mikrochir Plast Chir 35:72-82.
Fernandes M,Valente SG,Sabongi RG,Gomes Dos Santos JB,Leite VM,Ulrich H,Nery AA,da Silva Fernandes MJ(2018)Bone marrow-derived mesenchymal stem cells versus adipose-derived mesenchymal stem cells for peripheral nerve regeneration.Neural Regen Res 13:100-104.
Heidenreich A,Bellmunt J,Bolla M,Joniau S,Mason M,Matveev V,Mottet N,Schmid HP,van der Kwast T,Wiegel T,Zattoni F,European Association of Urology(2011)EAU guidelines on prostate cancer.Part 1:screening,diagnosis,and treatment of clinically localised disease.Eur Urol 59:61-71.
Jang S,Cho HH,Cho YB,Park JS,Jeong HS(2010)Functional neural differentiation of human adipose tissue-derived stem cells using bFGF and forskolin.BMC Cell Biol 11:25.
Kato H,Minamizato H,Ohno H,Ohira Y,Izawa T(2019)Exercise ameliorates high-fat diet-induced impairment of differentiation of adipose-derived stem cells into neuron-like cells in rats.J Cell Physiol 234:1452-1460.
Lin G,Banie L,Ning H,Bella AJ,Lin CS,Lue TF(2009)Potential of adipose-derived stem cells for treatment of erectile dysfunction.J Sex Med 6Suppl 3:320-327.
Lin G,Albersen M,Harraz AM,Fandel TM,Garcia M,McGrath MH,Konety BR,Lue TF,Lin CS(2011)Cavernous nerve repair with allogenic adipose matrix and autologous adipose-derived stem cells.Urology77:1509.e1-8.
Lu P,Jones LL,Snyder EY,Tuszynski MH(2003)Neural stem cells constitutively secrete neurotrophic factors and promote extensive host axonal growth after spinal cord injury.Exp Neurol 181:115-129.
Madonna R,Geng YJ,De Caterina R(2009)Adipose tissue-derived stem cells:characterization and potential for cardiovascular repair.Arterioscler Thromb Vasc Biol 29:1723-1729.
Makdissy N,Haddad K,AlBacha JD,Chaker D,Ismail B,Azar A,Oreibi G,Ayoub D,Achkar I,Quilliot D,Fajloun Z(2018)Essential role of AT-P6AP2 enrichment in caveolae/lipid raft microdomains for the induction of neuronal differentiation of stem cells.Stem Cell Res Ther 9:132.
Nagasaki H,Shang Q,Suzuki T,Hashimoto H,Yoshimura T,Kondo TA,Ozaki T,Maruyama S,Jomori T,Oiso Y,Hamada Y(2011)Low-serum culture system improves the adipogenic ability of visceral adipose tissue-derived stromal cells.Cell Biol Int 35:559-568.
Patrick CW,Jr.(2000)Adipose tissue engineering:the future of breast and soft tissue reconstruction following tumor resection.Semin Surg Oncol19:302-311.
Qian DX,Zhang HT,Ma X,Jiang XD,Xu RX(2010)Comparison of the efficiencies of three neural induction protocols in human adipose stromal cells.Neurochem Res 35:572-579.
Safford KM,Safford SD,Gimble JM,Shetty AK,Rice HE(2004)Characterization of neuronal/glial differentiation of murine adipose-derived adult stromal cells.Exp Neurol 187:319-328.
Safford KM,Hicok KC,Safford SD,Halvorsen YD,Wilkison WO,Gimble JM,Rice HE(2002)Neurogenic differentiation of murine and human adipose-derived stromal cells.Biochem Biophys Res Commun 294:371-379.
Santiago LY,Clavijo-Alvarez J,Brayfield C,Rubin JP,Marra KG(2009)Delivery of adipose-derived precursor cells for peripheral nerve repair.Cell Transplant 18:145-158.
Sun Y,Jin K,Xie L,Childs J,Mao XO,Logvinova A,Greenberg DA(2003)VEGF-induced neuroprotection,neurogenesis,and angiogenesis after focal cerebral ischemia.J Clin Invest 111:1843-1851.
Walsh PC(1987)Radical prostatectomy,preservation of sexual function,cancer control.The controversy.Urol Clin North Am 14:663-673.
Walz J,Burnett AL,Costello AJ,Eastham JA,Graefen M,Guillonneau B,Menon M,Montorsi F,Myers RP,Rocco B,Villers A(2010)A critical analysis of the current knowledge of surgical anatomy related to optimization of cancer control and preservation of continence and erection in candidates for radical prostatectomy.Eur Urol 57:179-192.
Ying C,Hu W,Cheng B,Zheng X,Li S(2012)Neural differentiation of rat adipose-derived stem cells in vitro.Cell Mol Neurobiol 32:1255-1263.
Ying C,Yang M,Zheng X,Hu W,Wang X(2013)Effects of intracavernous injection of adipose-derived stem cells on cavernous nerve regeneration in a rat model.Cell Mol Neurobiol 33:233-240.
Zarei MH,Soodi M,Qasemian-Lemraski M,Jafarzadeh E,Taha MF(2016)Study of the chlorpyrifos neurotoxicity using neural differentiation of adipose tissue-derived stem cells.Environ Toxicol 31:1510-1519.
Zhang H,Yang R,Wang Z,Lin G,Lue TF,Lin CS(2011)Adipose tissue-derived stem cells secrete CXCL5 cytokine with neurotrophic effects on cavernous nerve regeneration.J Sex Med 8:437-446.
Zhang HT,Luo J,Sui LS,Ma X,Yan ZJ,Lin JH,Wang YS,Chen YZ,Jiang XD,Xu RX(2009)Effects of differentiated versus undifferentiated adipose tissue-derived stromal cell grafts on functional recovery after spinal cord contusion.Cell Mol Neurobiol 29:1283-1292.
Zhao ZK,Yu HL,Liu B,Wang H,Luo Q,Ding XG(2016)Antioxidative mechanism of Lycium barbarum polysaccharides promotes repair and regeneration following cavernous nerve injury.Neural Regen Res11:1312-1321.
Zheng Y,Huang C,Liu F,Lin H,Yang X,Zhang Z(2017)Comparison of the neuronal differentiation abilities of bone marrow derived and adipose tissuederived mesenchymal stem cells.Mol Med Rep 16:3877-3886.
Zhong B,Liu X,Wu W(2018)Transplantation of neural stem cells from different sources in the treatment of cerebral ischemia injury in rats.Zhongguo Zuzhi Gongcheng Yanjiu 22:3349-3356.