ABSTRACT Introduction Regenerative surgery is a branch of medicine that has as the main goal to regenerate cells, tissue and organs that are damaged. In this branch, adipose tissue has a central role, especially related to mesenchymal stem cells that derive from it. Studying cells and tissues, both in science and in medicine, cell counting is one of the most used procedures: it consists in a process in which a sample of cells is counted to determine the number of cells and its viability; This can be done both manually and with automatic cell counters. Viability is the number of live cells over the total number of cells of the sample. The most used way to determine viability is by using Trypan Blue, also used in this study. Materials and method In this initial study LUNA-II TM by Logos Biosystem has been used inside ORs to study different samples of adipose tissue. 17 samples of adipose tissue, from different patients, with different conditions and from different parts of the body, have been analyzed. Goals This initial study has as its main goal to study if an automated cell counter can be used inside an OR and if this machine can represent a good perspective for Regenerative Surgery. Secondly the goals are to study the difference of viability of adipose tissue to see if it is possible to have a new perspective of research fields. Discussion LUNA-II TM has been used to analyze 17 samples as an intra-operatory machine. Thanks to its small dimension and the fact that it is very easy to use, it can easily fit the OR, the only thing to remember is that it needs to be plugged, and it is important to remember to do it in a safe way for all the members of the team. LUNA-II TM and the procedure to count cells, are easy and intuitive, so it is possible to train the person responsible for the count easily and fast. The viability of adipose tissue resulted 77,76% as average of the viability, and 78,00% as median of the viability. According to the preliminary data it seems that patients with MSL1 have lower adipose tissue viability, also it seems that patients who had suffered from obesity in the past have a very little lower viability compared to healthy patients. Regarding differences of viability connected with different samples from different body parts, it seems there is no difference except for the inner knee, that showed higher viability, and lumbar region, that seems to have lower viability. The limitation of this study is the number of samples to evaluate differences in viability regarding diseases and different body parts. Conclusions LUNA-II TM it’s a very simple machine, thanks to its intuitiveness, fastness and the fact that it is very reliable, it can be used inside an OR. Thanks to its fastness it can be used both for intraoperative procedures and for research purposes. According to our results, adipose tissue’s viability is 70-80%, and it seems that there might be differences between different pathologies of adipose tissue and body parts, but this needs to be explored with more samples. Cell counter seems to be an important resource for Regenerative Surgery and can become an important feature to the OR procedures involving adipose tissue and also be important for the progression of the research and knowledge. Procedures that might take advantage from this machine are lipofilling and MSCs implant.
Introduzione La Chirurgia Rigenerativa è una branca della medicina che si pone l’obiettivo di rigenerare cellule, tessuti ed organi danneggiati. In essa gioca un ruolo centrale lo studio del tessuto adiposo. Nello studio di cellule, una delle procedure più utilizzate è la conta cellulare: valuta il numero di cellule del campione e contestualmente, è possibile valutare la viability, ossia il numero di cellule vive sul totale delle cellule del campione. Per farlo si possono usare vari metodi di colorazione, il più usato è il Trypan blue, applicato anche in questo studio. Materiali e metodi In questo studio preliminare è stato utilizzato un cell counter autonomizzato, LUNA-II TM, slide per la preparazione del campione e Trypan Blue. Sono stati raccolti 17 campioni, con diverse caratteristiche in merito a patologie del tessuto adiposo, e provenienti da diverse zone del corpo. Obiettivi Questo studio preliminare si pone come obiettivo principale studiare la fattibilità d’uso del cell counter come strumento intraoperatorio e in chirurgia rigenerativa, secondariamente, studiare la viability del tessuto adiposo. Discussione Il LUNA-II TM è stato impiegato per l’analisi di 17 campioni come strumento intraoperatorio; può essere facilmente inserito all’interno della sala operatoria, l’unico vincolo è la necessità di essere collegato ad una presa di corrente. Data la semplicità d’uso, è possibile istruire in modo rapido ed efficace l’operatore addetto alla conta. La viability del tessuto adiposo sottocutaneo prelevato mediante lipoaspirazione risulta essere in media 77,75%, con mediana 78,00% e sembra risultare che il tessuto adiposo dei pazienti con LMS 1 presenti una viability minore, mentre nei pazienti che hanno sofferto di obesità, la viability sembra di poco minore rispetto ai soggetti sani. Per quanto concerne invece le differenti viability zona-specifica, i risultati preliminari sembrano indicare una minore viability della zona lombare, e una maggiore viability della zona dell’interno ginocchio. Il limite di questo studio è la numerosità dei campioni pertanto. Conclusioni Il cell counter LUNA-II TM risulta uno strumento che può essere utilizzato all’interno della sala operatoria. Secondo i risultati preliminari, la viability del tessuto adiposo prelevato mediante lipoaspirazione è compresa tra il 70-80%, sembra essere significativamente inferiore nei pazienti affetti da LMS1 ed essere di poco più bassa nei pazienti post-bariatrici. La zona lombare sembra avere una viability inferiore, mentre la zona dell’interno ginocchio sembra avere una viability maggiore, rispetto alle altre zone prese in analisi. Il cell counter sembra dunque essere una risorsa importante per la chirurgia rigenerativa. In particolare la tecnica di lipofilling e l’impianto di MSCs derivate da tessuto adiposo sembrano essere le procedure che potrebbero trarre i maggiori vantaggi dall’uso di questo dispositivo.
"Cell Counting": nuovo approccio clinico sperimentale alla Chirurgia Rigenerativa
GAGGIATO, GRETA
2021/2022
Abstract
ABSTRACT Introduction Regenerative surgery is a branch of medicine that has as the main goal to regenerate cells, tissue and organs that are damaged. In this branch, adipose tissue has a central role, especially related to mesenchymal stem cells that derive from it. Studying cells and tissues, both in science and in medicine, cell counting is one of the most used procedures: it consists in a process in which a sample of cells is counted to determine the number of cells and its viability; This can be done both manually and with automatic cell counters. Viability is the number of live cells over the total number of cells of the sample. The most used way to determine viability is by using Trypan Blue, also used in this study. Materials and method In this initial study LUNA-II TM by Logos Biosystem has been used inside ORs to study different samples of adipose tissue. 17 samples of adipose tissue, from different patients, with different conditions and from different parts of the body, have been analyzed. Goals This initial study has as its main goal to study if an automated cell counter can be used inside an OR and if this machine can represent a good perspective for Regenerative Surgery. Secondly the goals are to study the difference of viability of adipose tissue to see if it is possible to have a new perspective of research fields. Discussion LUNA-II TM has been used to analyze 17 samples as an intra-operatory machine. Thanks to its small dimension and the fact that it is very easy to use, it can easily fit the OR, the only thing to remember is that it needs to be plugged, and it is important to remember to do it in a safe way for all the members of the team. LUNA-II TM and the procedure to count cells, are easy and intuitive, so it is possible to train the person responsible for the count easily and fast. The viability of adipose tissue resulted 77,76% as average of the viability, and 78,00% as median of the viability. According to the preliminary data it seems that patients with MSL1 have lower adipose tissue viability, also it seems that patients who had suffered from obesity in the past have a very little lower viability compared to healthy patients. Regarding differences of viability connected with different samples from different body parts, it seems there is no difference except for the inner knee, that showed higher viability, and lumbar region, that seems to have lower viability. The limitation of this study is the number of samples to evaluate differences in viability regarding diseases and different body parts. Conclusions LUNA-II TM it’s a very simple machine, thanks to its intuitiveness, fastness and the fact that it is very reliable, it can be used inside an OR. Thanks to its fastness it can be used both for intraoperative procedures and for research purposes. According to our results, adipose tissue’s viability is 70-80%, and it seems that there might be differences between different pathologies of adipose tissue and body parts, but this needs to be explored with more samples. Cell counter seems to be an important resource for Regenerative Surgery and can become an important feature to the OR procedures involving adipose tissue and also be important for the progression of the research and knowledge. Procedures that might take advantage from this machine are lipofilling and MSCs implant.| File | Dimensione | Formato | |
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https://hdl.handle.net/20.500.12608/30543