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Assessment of Biological Properties of Bone Marrow Mesenchymal Stromal Cell by Ex-Vivo Expansion and Differentiation

Thesis Info

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Author

Satti, Humayoon Shafique

Supervisor

Salman A Malik

Program

PhD

Institute

Quaid-I-Azam University

City

Islamabad

Province

Islamabad.

Country

Pakistan

Thesis Completing Year

2017

Thesis Completion Status

Completed

Subject

Biochemistry

Language

English

Link

http://prr.hec.gov.pk/jspui/bitstream/123456789/9799/1/Humayoon%20Shafique%20Satti_Biochem_2017_QAU_Main%20part.pdf

Added

2021-02-17 19:49:13

Modified

2024-03-24 20:25:49

ARI ID

1676725537526

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Mesenchymal stromal cells (MSCs) represent a rare population of multipotent stem cells found in many adult tissues. In bone marrow MSCs support haematopoiesis by providing marrow stroma. MSCs have recently emerged as an exciting candidate for cellular therapy due to their hypoimmunogenic properties and ability to differentiate into different tissue types. With the intent of optimizing large-scale expansion of MSCs for clinical use, we compared different culturing conditions for their ability to support growth and proliferation of bone marrow MSCs (BM-MSCs). We evaluated cells cultured in Alpha Modified Eagle Medium (αMEM), Dulbecco’s modified Eagle medium (DMEM) in the presence of either 10% fetal bovine serum (10% FBS), 10% pooled human platelet lysate (10% pHPL) or commercial serum-free formulation (StemPRO® MSC SFM). We concluded that supplementing growth medium with pHPL resulted in superior cell yield than fetal bovine serum (FBS) and comparable to commercial serum-free formulation. Study II was a phase-I human clinical trial, establishing safety of autologous BM-MSC transplantation in nine spinal cord injury patients. For this ex vivo expanded MSCs were injected intrathecally after premedication. All of our patients tolerated the procedure well and no complication was observed during a median follow up of 644 days. Some of the patients with sub-acute disease reported subjective improvement in sensory and neurological functions. Lastly we carried out a phase-I/II trial of MSCs in allogeneic use as a treatment option in treating steroid-resistant graft versus host disease (GVHD). GVHD is a life threatening complication of allogeneic stem cell xii transplantation and only a fraction of patients are cured through steroids. Those who fail to respond to steroid have a very poor prognosis. The patients received third party BM MSCs as their bone marrow MSCs were failing to cope with the immunemediated tissue destruction by donor T cells. A total of 33 MSC infusions were given to 10 patients suffering from acute GVHD (n=3), chronic GVHD (n=5) and overlap syndrome (n=2). Eight out of ten patients are alive after a median follow up of 11 months with five having sustained CR. Three patients with partial response received further doses to sustain response. One of them died of lung infection while others are still alive. One out of the two non-responding patients died after 17 days of MSC therapy due to advanced liver GVHD while the other is having stable disease course. The overall and disease-free survival was 80% and 50% respectively. The patients developed no complication or toxicity related to MSC infusion. Luminex analysis revealed a modest drop after MSC infusion in pro-inflammatory cytokines like IFNγ, TNFα, IL-1β, IL-2, IL-4, IL-6, IL-17A and IL-17F, whereas serum IL-10 levels were slightly raised. Overall a total of 57 preparations of cultured stem cells were made which were transplanted via I.V. or I.T. route with no adverse event or serious complication to report. Therefore it is concluded that MSCs can be safely given in both autologous as well as allogeneic setting as cellular therapy in selected clinical situations. This initial data on safety and efficacy calls for taking these clinical trials into advanced phase with more detailed account of efficacy measures and addition of placebo groups.
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قاضی احمد میاں اختر جونا گڑھی

احمد میاں اختر جونا گڑھی
ہندوستان اور پاکستان کی علمی دنیا کا یہ بڑا افسوس ناک سانحہ ہے کہ گزشتہ مہینہ مشہورصاحب علم و قلم قاضی احمد میاں اختر جونا گڑھی نے کراچی میں انتقال کیا، وہ اسلامی علوم کے ممتاز فاضل تھے، خصوصاً اسلامی تاریخ پر ان کی نظر نہایت وسیع تھی اور ان کا علمی و تحقیقی مذاق بہت بلند تھا، عربی، انگریزی اور اردو تینوں زبانوں میں یکساں دستگاہ حاصل تھی اور تینوں میں مضامین لکھتے تھے، ان کے مضامین ہندوستان و پاکستان کے تمام سنجیدہ علمی رسالوں میں نکلتے تھے، معارف کے پرانے مضمون نگارتھے، ان کے انگریزی مضامین کا ایک مجموعہ شیخ محمد اشرف تاجر کتب لاہور نے شائع کیا ہے، اقبال پر ان کی ایک کتاب حال ہی میں چھپی ہے، دارالمصنفین سے بھی ان کی ایک کتاب ابن صاعد اندلسی کی طبقات الامم کا ترجمہ عرصہ ہوا شائع ہوچکی ہے، اگر ان کے مضامین جمع کیئے جائیں تو کئی جلدوں میں آئیں گے۔
مرحوم ریاست جونا گڑھ کے جاگیردار تھے، وہاں کے انقلاب میں لٹ لٹاکر بڑی مصیبتوں سے کراچی پہنچے، کچھ دنوں تک انجمن ترقی اردو سے وابستہ رہے، اس کے بعد سندھ یونیورسٹی میں شعبہ اسلامیات کے صدر ہوگئے تھے، طبعاً نہایت شریف، متواضع اور خاکسار تھے، دارالمصنفین سے ان کا تعلق بڑا پرانا اور مخلصانہ تھا ابھی انھوں نے پاکستان میں دارالمصنفین کی کتابوں کے لئے لائسنس دلانے میں بڑی مدد کی تھی، اب اس زمانہ میں ایسے صاحب کمال کی جگہ کابھرنا مشکل ہے اﷲ تعالیٰ اس شیدائے علم کو اپنی رحمت و مغفرت سے سرفراز فرمائے، انتقال کے وقت ۶۰ سال سے اوپر کی عمر رہی ہوگی۔ (شاہ معین الدین ندوی، ستمبر ۱۹۵۵ء)

 

Role of Religious Interventions in the Reintegration of Prisoners: A Case Study of Selected Jails of Khyber Pakhtunkhwa, Pakistan

Prisoners’ reintegration is the core concept of almost all penal systems in the world. One of the potent tools to ensure prisoners’ reintegration is effective network of religious services within prisons. This paper aims at exploring the role of religious interventions in the reintegration of prisoners with specific focus on Khyber-Pakhtunkhwa (KP) jails.  Six high profile jails---Central Jail Peshawar, Haripur, Bannu, and District Jail Timergara, Mardan and Kohat of KP were purposively selected. Mixed methodology, more specifically concurrent triangulation technique, was used to collect and analyze the data. Of all 261 respondents, 250 comprised of jail inmates (under-trial and convicted adults and juveniles male prisoners) were randomly selected within the six jails of the province and interviewed through semi-structured questionnaire. The remaining 11 respondents, purposively selected and interviewed through interview-guide included jail officials of all the selected prisons (6 in numbers) and ex-prisoners (5 in numbers). It was found that a clear majority of the respondents considered religious interventions instrumental in accomplishing the goal of prisoners’ reintegration i.e, making them law abiding, productive, contributing and pro-social citizens. Many of the apparently incorrigible and potentially dangerous prisoners altered the course of their lives once they went through religious programs inside prisons. It was also discovered that in KP prisons, there was no effective network of chaplaincy services, and often these services were provided by self-motivated religious prisoners and rarely by the prison management with the collaboration of NGOs.  Yet, the existing religious interventions had an extraordinary impact in terms of reforming the inmates. Hence, it is recommended that any prisons’ reform strategy must incorporate a well-designed framework of religious programs to transform criminals into an asset for society.

Comparative Sorption Studies of Divalent Metal Ions on Nickel Oxide Nio

The present work focuses on the characterization, potentiometric titration, kinetics and adsorption studies of a novel and efficient adsorbent (NiO). The surface structure of the NiO was identified and characterized by X-ray diffractometry (XRD), FTIR spectroscopy, surface area, point of zero charge (PZC), transmission electron microscopy (TEM), thermogravimetric and differential thermal analyses (TGA/DTA), scanning electron microscopy (SEM) coupled with energy dispersive X-ray analyses (EDX). The dissolution study of NiO was undertaken as a function of pH (2.00-11.00) and temperature (283-323K), which was observed to increase substantially by decreasing the initial pH of the system. The point of zero charge (PZC) of NiO in the presence of background electrolyte was determined by the salt addition, mass and fast titration techniques in the temperature range 303-333 ± 1K. The PZC of NiO determined by salt addition method was comparable in magnitude to the one obtained by mass titration technique. The coincidence between the PZC and CIP indicated that no specific adsorption of the electrolyte occurs at pHPZC of NiO and the surface carries a net zero charge at CIP. However, a shift in the PZC values with concentration and nature of divalent metal cations indicated their specific adsorption onto NiO surface. The PZC and the concentration of NiOH2+ groups were decreased whereas the concentration of NiO- was increased with the increase in temperature of the medium. A comprehensive study of the potentiometric titrations of NiO in the presence of Na+, Cd2+, Zn2+, Ni2+, Pb2+, Cu2+, Ca2+, and Mg2+ was conducted under different experimental conditions of temperature, concentration and pH. The affinity of metal cations evaluated from the potentiometric titration data was found to be in order: divalent transition metals > alkaline earth metals > alkali metal. The deprotonation of NiO was observed to be dependent upon the concentration, temperature of metal cations present in the system. The changes in enthalpy (ΔH) and entropy (ΔS) connected with the surface deprotonation of the NiO/electrolyte interface were measured. The loss in entropy indicated that potential determining ions were less hydrated at the interfacial region. Thermodynamic parameters showed that the positive value of enthalpy was the driving force for the deprotonation of the nickel oxide surface. The kinetics of metal cations sorption onto NiO were studied at different temperatures (303–323K). The applicability of the various kinetic models was checked to determine the mechanism of adsorption. The pseudo first order model was best fitted among the kinetic models to describe the kinetic data. The values of activation energy (Ea) determined from Arrhenius equation were observed to be 29.40, 43.74, 32.66, 3.77 and 12.96 kJ mol-1 for Cd2+, Zn2+, Co2+, Pb2+ and Ca2+ respectively. The cation exchange sorption of Cd2+, Zn2+, Pb2+, Co2+, Ca2+, and Mg2+ from aqueous solution on NiO was studied under batch process as a function of concentration of metals, amount of adsorbent and temperature of the suspension. The effect of initial solution pH on divalent metals removal from aqueous solution was examined to assess the sorption behaviour of NiO at different pH values. The solution pH was found to play a decisive role in the metal ions precipitation, surface dissolution and adsorption of metal ions onto the NiO. The preferential uptake of divalent metals from their co–ions was observed in the order: NO3- > Cl- > SO42- which reflected that the presence of nitrate ions was more effective in exchanging the adsorption of divalent metals as compared to chloride and sulfate anions. Desorption of divalent metals from impregnated NiO was checked with protons by varying the initial pH of the suspension at 303 ± 1 K. The adsorption experiments showed that the selectivity of NiO towards different divalent metals followed the trend: Pb2+ > Zn2+ > Co2+ > Cd2+ > Ca2+ > Mg2+ which was in a line with the first hydrolysis constant values of these metal cations. Korbatov equation was used successfully to derive the H+/M2+ stiochiometry of the ion exchange reaction. The exchange between the proton from the NiO surface and the metal from solution was responsible for the adsorption. The uptake of divalent metals by NiO was achieved neither by the replacement of Ni from NiO nor due to precipitation of metal at pH 7.50. The cation exchange data was explained with the help of Freundlich and Langmuir models. The isosteric heat of adsorption (ΔΉ) was also calculated between 303-323 K. The Langmuir constants were used to compute the apparent thermodynamic parameters DH, DS and DG. The positive DS values demonstrated that the adsorption reaction was spontaneous. The spontaneity of the metals adsorption onto the NiO was justified thermodynamically by the decrease in Gibbs free energy. The increase in the entropy (DS) of the system followed the trend: Co2+ > Ca2+ > Zn2+ > Pb2+ > Cd2+ > Mg2+ which was almost parallel to their corresponding enthalpic values. The cation exchange sorption of divalent metals by the NiO was endothermic driven by entropy increase in the system. The spectroscopic analyses of the solid media also give a strong support to the conclusion that divalent metal ions were chemisorbed onto the surface of nickel oxide.