Factors Affecting Implantation Rate in Embryo Glue Application - Literature review Example

Embryo Glue: A Critical Literature Review Contents Introduction 3 Physiology of Implantation 3 Viscosity and Culture Media 5 Application in Animals and Humans 7 Factors Affecting Implantation Rate in IVF Processes 9 Physiology of Implantation 10 Clinical Data of Patient Characteristics & IVF 12 Research Evidence on Improving Implantation 13 Timing and Risk Factors in Embryo Glue Application 14 Conclusion 16 Bibliography 17 Introduction Assisted-Reproduction involves the use of various technologies to cause fertilisation and foetal implantation in the womb of a patient (Kennedy, 1997). One of the approaches is In-Vitro Fertilisation (IVF) where the fertilisation of the egg and the sperm occurs outside the body and is introduced to the female body (Schoolcraft, et al., 2001). This includes the use of various physiological techniques to take the relevant cells of the two parties, cause the fertilisation and introduce it to the female body through a culture media. “Bioadhesive” is a broad term that refers to a set of biological monomers that enable cells to be attached to various tissues in living things (Ulkoski & Scholz, 2014; Pande, et al., 2014). This term has evolved to describe a form of glue that is created from these monomers which usually contain proteins and carbohydrates. Bioadhesives have numerous practical applications in medical science, most notably, in Embryo Glues which is used as an aid for conception in humans. Embryo glues are concentrates that aid implantation in various external fertility processes, mainly IVF (Tomari, et al., 2014). The purpose of this document is to critically review and evaluate important features of the physiology of implantation and improvement of implantation in IVF with an emphasis on Embryo Glue as a part of the IVF process. To this end, the paper will critique the physiology of embryo glue and its inherent features when applied in practice. It will go further to critique the factors that affect the success rate of embryo glue. Finally, timing and other risk factors affecting the successful implantation of embryos will be discussed in this paper. Physiology of Implantation The entire process of carrying out IVF involves the selection of the most appropriate cells from the sperm and egg of the patient in order to carry out the fertilisation externally (Schoolcraft, et al., 2001). When this is done, the next step involves the introduction to the body which is contingent on the circumstances and features of the recipient. The normal features of the average recipient in the different parts of the female reproductive system include: Oviduct Uterus Glucose concentration 0.5mm 3.15mm pH 7.5 7.1 Oxygen concentration 8% 1.5% Carbon dioxide 12% 10% Glycine concentration 2.77% 19.33% Statistics of the Features of the Female Reproductive System (Gardner & Lane, 2007) Due to the nature of the oviduct and uterus, there is the need to deposit embryo 2cm below the uterus fundus (Aflatoonian & Asgharnia, 2006). This allows the embryo to move and survive the conditions within the uterus and oviduct. Since the environments change between the two organs, there is the need for a carefully composed culture media to be created to ensure the safety of the fertilised egg. The main variables that are varied and important to ensure the survival of the fertilised egg includes ensuring that the Ion/Water channels are appropriate and there are no blockages to implantations within these systems (Gardner & Lane, 2007). There is often the need to conduct a trial transfer to check environmental conditions in the organs and also achieve blastocyst activation (Aflatoonian & Asgharnia, 2006; Zhang, et al., 2013). There is the need to ensure that the fundus are clean and there is nothing affecting it in any way (Aflatoonian & Asgharnia, 2006). In order to achieve blastocyst activation, it is advisable that there are amino acids that are cultured in the media for 48 hours to 93 hours to ensure that the there is sufficient nourishment to keep the fertilised egg active (Lane & Gardner, 1994). Embryo glue contains saccharides and amino acids that are meant to create an environment and culture media through which the fertilised embryo can move into the womb without any blockages or inhibitions (Tomari, et al., 2014). Embryo glue also has a relatively high concentration of hyaluronan which is an aid in causing the fertilized egg to attach itself to the walls of the womb to commence the process of nourishment and dependence on the mother (Tomari, et al., 2014). It also contains recombinant albumin which is the most basic form of protein that an embryo can depend upon until it is attached to a womb and can develop a nourishment relationship with the system of the mother (Nakagawa, et al., 2012). Viscosity and Culture Media The ideal viscosity rate for a culture media is 200gm at 5ml of a pellet of fresh medium and this allows for the creation of an optimal density that allows for the movement of the embryo (Brinsden, 2010). This is because an extremely thick culture media might hinder the progression of the embryo. A low dense media might not contain all the appropriate nutrients and nourishments that might be necessary for a successful IVF. In order to achieve insemination, there is the need for an appropriate ion/water channel, oestrogen or progesterone, as well as proteins acting as stimulator (Liu, et al., 2014). There are other necessary components like glucose, vitamins and cholesterol that must be mixed in the right proportions to ensure the culture media works appropriately (Xella, et al., 2010). Albumen is the main macromolecule in most embryonic culture media and it provides the appropriate level of nourishment for a fertilised egg (Tomari, et al., 2014). Serum albumin is derived from blood and due to that fact, it carries with it the risk of contamination from viruses and other micro-substances (Bontekoe, et al., 2013). However, studies show that hyaluronic acid can be an appropriate replacement for albumen as the main macromolecule in the embryo transfer medium and this can be presented in a form that is properly monitored to ensure that they do not contain viruses or other harmful substances (Simon, et al., 2003). The chemical composition of the culture media that is created in Embryo Glue includes Anionic, non-sulfated glycosaminoglycan and hydrauluronic acid which is a naturally occurring component in most vertebrate mammals (Furnus, et al., 1998). These three ingredients are added to the culture media in three to five days to change the component of a fertilised egg in IVF. The primary beneficial content of the Embryo Glue is the hyaluronic acid which naturally occurs as a molecule and is naturally present in the female reproductive tract as a macromolecule (Salamonsen, et al., 2001). This creates a mechanism through which implantation can be enhanced. It is demonstrated from previous research that hyaluronic acid increases cell-to-cell adhesion and cell-to-matrix adhesion (Bontekoe, et al., 2013). Hyaluronic acid increases the viscosity of the mixture to levels similar to traditional fluids in the uterine and this is due to the biological features of the hyaluronic acid which comes with the anionic, nonsulfated glycosaminoglycan components (Furnus, et al., 1998). The thickened fluid promotes transfer and motion of the embryo and also prevents the expulsion or backward movement of the embryo through diffusion (Simon, et al., 2003). Most scientists seem to agree on the fact that the viscosity of the Embryo Glue helps to aid motion and implantation. However, some state that viscosity is not the only reason for this capability of the embryonic glue, since there are numerous viscous gels that do not have the capability to promote onward movement of cells and aid adhesion to other cells. The human embryo also contains CD44 receptors which enables the human embryo to bond with other cells until the blastocyst is formed, which commences the gestation process (Nakagawa, et al., 2012). Nakagawa et al posit that hyaluronic acid activates the competencies of CD44 and ensure that they are most active when applied through the Embryo Glue. This enhances and optimises the embryo transfer media and makes it more potent and improves efficiency of external fertilisations leading to actual conception. The next most important ingredient in the Embryonic Glue is the fibrin which influences the transfer media and plays a major role in the culture media of EG. Fibrin is made up of fibrinogen and thrombin which is introduced into the Embryonic Glue with an aprontinin (Bontekoe, et al., 2013). Fibrin, which is a viscous substance it acts as a sealant and enables the embryo to firmly adhere to the tissue (Bontekoe, et al., 2013). This implies that it reduces the risks related to embryo expulsion. Another benefit it has is that it prevents ectopic pregnancy and ensures that the embryo sticks to the womb and not other inappropriate parts of the female reproductive system. Therefore, as long as the embryo remains as a clot, the fibrin will prevent it from being expelled. And this occurs until the clot dissolves. Application in Animals and Humans IVF has been applied to various cases in animals for research and other purposes. One of the most significant findings indicate that the best results of IVF is to introduce the fertilised egg into the female body of rodents when less than 35% of the cells are formed (therefore, introducing it where the cell formation process is still not completed) and also introducing it 14 hours less than the normal maturation process (Sjoblom, et al., 1999). This actually increases chances of implantation by over 50% and involves reduced viscosity in the culture media and a more active culture media (Sjoblom, et al., 1999). The application of Embryo Glue has been tested in animals and this complements the study of how it works in human beings. Findings from some studies indicate the fact that hyaluronic acid’s efficiency and effectiveness is strongly influenced by the nature of the receptors of the species of animal on which it is being applied. Figure 1: Kinetic Parameters of Hyaluronic Acid Application in Animals (Necas, et al., 2008) The study shows that different species have extraction ratios and different parts of the body on which hyaluronic acid (HA) is applied leads to different resources. However, most of the situations where it is applied indicate that it is applied in the form of plasmic and viscous forms, like our case of interest, Embryonic Glue. Figure 1 above shows that the ratio for hepatic application in pigs (hepatic) and sheep are quite high, 50%. This culminates in high levels of plasma clearance each day and high total daily turnover. This provides a general context and a general framework within which Embryonic Glue’s application in animals can be estimated or perceived. And it is observed that the “biocompatibility, biodegradability, and chemical structure of hyaluronic acid must be modified to meet the needs of different species of animals for optimum results” (Necas, et al., 2008, p. 408). Early studies in animals showed that urinary contractions have an effect on implantation in embryos and the utilisation of Embryo Glue (Madani & Jahangiri, 2013). This was not openly detected in humans and the evidence was marginal and could only be seen through digitalised images (Schoolcraft, et al., 2001). Studies show that bigger mammals have bigger success rates of applying HA and this is evidenced from proceedings in the cattle insemination industry (Babiuk, et al., 2012). The natural structures of animals play a role in the success or failure rate of Embryo Glue application as it determines how well movement might occur and how well embryos can attach to wombs of the respective species (Figard & Sokac, 2011). Therefore the unique features of each creature determines the kinetics and the receptivity of Embryo Glue and this gives room for additions and dosage which will alter the viscosity of the appropriate dose for each species. For instance, mice have a very high rate of implantation on the same day and require less HA dosage (Babiuk, et al., 2012). Factors Affecting Implantation Rate in IVF Processes Evidence indicates that there is a slight increase in the chances of implantation when Embryo Glue is applied in an external fertilisation process, albeit a very small increase in probability of about 3% (Fancsovits, et al., 2014). This implies that the implantation rates are influenced and induced by some kind of physiological factors and pointers which make the Embryo Glue a better and a stronger option. Further research by other scientists indicates that in fertile couples, Embryo Glue does not really make any changes and improvement to the implantation rates (Safari, et al., 2014). This implies that the assistive nature of Embryo Glue only extends to people whose fertilised eggs need some kind of support and assistance in pharmacokinetic aspects and areas like adhesion and fusion. These are things the normal natural reproductive system can achieve without need for any kind of assistance. This leads to the discussion of the actual physiology of implantation in the womb. Although much study ought to be done to prove some important pointers, some pointers have been logically presented to be representative of the main elements and aspects of implantation success rates. The physiological features of the parents who are going through the process is very important and crucial. This is because the parents’ genes determine and define the way through which the foetus will be accepted. And this will also determine the right dosage that can be applied to achieve the proper results for the parents. Therefore, there is the need for a doctor to check the profile and history of the patients and make appropriate recommendations in order to achieve optimal results in every aspect of the insemination and the process. Irregular-shaped bladders and abnormal bladder pressures could make it difficult for the implantation to work as they interfere with the movement of the fertilised egg during the introduction through the fertility duct of a patient (Atala, 2012). A filled bladder simplifies the embryo transfer process and makes it much easier for the movement to occur during the process (Madani & Jahangiri, 2012). Physiology of Implantation Implantation without Embryo Glue comes with numerous results and functions. The normal medium like Universal/IVF Media is a replication of the luteal phase which includes oestrogen and progesterone (Xella, et al., 2010; Zhang, et al., 2013). These contain some natural cells like Granulosa cells that work to protect the fertilised egg from damages and blockages (Kim, et al., 2013). However, evidence shows that there is no clear indication that the use of natural and alterantive culture media that does not utilise Embryo Glue improves or reduces the results of implnatation (VerMilyea, et al., n.d.). However, other studies show that there are higher levels of results of about 10% difference in couples who used other culture media in achieving implantation (Mahani & Davar, 2007). This is mainly due to the receptivity and attachment element of hyaluronic acid as a bioadhesive which leads to early cleavage (Gardner & Lane, 2007; Zhang, et al., 2013). Pure research into Embryo Glue shows that the major content, Hyaluronic acid (HA) is a glycosaminoglycan which is a component of extracellular matrix which occurs in teh female reproductive duct (Simon, et al., 2003). Thus, the utilisation of HA creates some kind of supplement that contains enhanced features that improves development in the fertilised embryo from the time it is introduced into the culture media to the blastocyst stage in order to maintain and retain embryo’s natural functionalities after the embryo is moved from a frozen state. Due to this, the highest levels of implantations are reported in situation where the embryo transfer was done after the introduction of HA as the main macromolecule in the culture media since it provides extra safeguards and processes that allows implantation to occur (Gardner, et al., 1999). There are different conflicting views of the actual assistance and support HA gives to the concept process when the Embryo Glue is applied. The fundamental physiological benefit of HA is the fact that it mediates the interactions between the embryo and the surface of the endometrium which is through the adhesion of the embryo with the womb. However, other studies indicate that it acts as an autocrine medium that stimulates the cells within the reproductive duct to perform their functions as required (Furnus, et al., 1998). This is because it induces actions that accelerate the other fluids in the reproduction system to work and achieve the required ends in order to secure a pregnancy. However, Gardner et al, who studied the processes of HA and how it works in Embryo Glue came up with another thesis which asserts that the highest number of cells and implantation levels occurred when HA was combined with serum albumin in the same medium to ensure high nutritional levels (1999). This is because there is a higher probability and a stronger sense of nutrition and nourishment for the fertilised egg when there is serum albumin which provides proteins and other proteins for improved and enhanced mediums. Therefore, it can be said that the content and the physiology of a given brand of Embryo Glue provides the main deciding factor that determines the success of a given implantation process. Clinical Data of Patient Characteristics & IVF The presence of barriers and inappropriate substances in the female reproductive system of a patient plays a major role in the attainment of success in IVF (Liu, et al., 2014). There are some inherent elements and processes of the recipient which influences the way and manner within which IVF works and achieves results. Some of the pointers include: 1. Ovarian reserve (fresh ovaries are less successful in implantation than frozen ovaries) 2. Hormonal levels 3. Age (Al-Azawi, et al., 2012). The receptivity of the endometrium system of a patient is significant in ensuring the success or failure of an IVF transfer since it determines interface synchronisation (Kuc, 2012). Ideally, an endometrial thickness of 4mm – 7mm achieves optimal results in IVF (Garcia-Velasco, n.d.). The hormonal levels relate to the oestrogen quality and catchotestrogen features and quality (Paria, et al., 2001). These features also determine the quality of the embryo to withstand excess conditions in the culture media and diseases that might stand in the way of an embryo. Most patients who are considered mainstream in IVF are patients aged below 40 (Coughlan, et al., 2014). Patients above 40 are considered patients with biological features that are not so positive or helpful with IVF. Other issues like sperm DNA fragmentation and fibroids and endometrium polyps play a role in determining whether a patient’s implantation rates will be high or low (Coughlan, et al., 2014). It is identified that Embryo Glue is most appropriate for infertile patients as there is little change in normal patients (Valojerdi, et al., 2006). There is also additional evidence to the effect that Embryo Glue and the HA content have a stronger impact on persons who have gone through several unsuccessful implantation attempts (Nakagawa, et al., 2012). This leads to a 41% chance of conception in women of the same age who are infertile as opposed to 35% for those who used natural means of insemination (Nakagawa, et al., 2012). This is because such patients are able to get higher results of successful conception, especially, after they are able to go through a renewal of their hormones and changes in the situations they go through. This implies that Embryo Glue is more of a cure to unsuccessful implantation. There have been deeper insights into the different features and it has been identified that the age and timing of the insemination is very important and significant in determining whether the application of Embryonic Glue will lead to implantations or not (Figard & Sokac, 2011). It is observed that there linings of the wombs of older women (40+) are harder and less receptive of the fertilised embryo (Figard & Sokac, 2011). This means that the use of HA in attempting to attach a fertilised egg into an older womb has a lower probability of resulting in conception than the case might be for younger women who have a more receptive womb lining. Research Evidence on Improving Implantation There are several pointers that have come to the fore that shows the extent to which changes must be conducted in order to achieve results in implantation. The most basic pointer for achieving successful implantation in the womb is the use of the appropriate catheter type and the careful placing of the embryo (Boomsa, 2006). Aflatoonian and Ashgamia (2006) identify that careful placement of the embryo involves placing it just 2cm below the uterus fundus. This leads to an optimal functioning of the culture media and enhances embryo function and gives a greater chance of implantation. Studies also show that there is the need for the evaluation of cervico-uterine axis and performance in a mock transfer and the careful evaluation of the uterine capacity (Madani & Jahangiri, 2013). There is the need for cervical preparation by removing mucus or blood on the catheter before loading the embryo medium (Madani & Jahangiri, 2012). It is also identified that after introducing the embryo and culture media, there is the need for the prevention of the expulsion of fluids with the use of an appropriate sealant (usually a fibrin sealant) as well as appropriate bed rest for the patient (Madani & Jahangiri, 2012). Implantation has been viewed as more appropriate and more likely when the patient’s condition is thoroughly examined and the dosage of the mixture is varied appropriately to meet specific needs of the patient (Simon, et al., 2003). This includes the introduction of amino acids and other forms of gases like oxygen in the right proportions to guarantee optimal performance (Gardner & Lane, 2007). The techniques and methods of utilisation must also be varied as well as the grading of the quality of the egg to achieve optimal results (Fancsovits, et al., 2014). Timing and Risk Factors in Embryo Glue Application This section of the literature review will critically examine and review aspects and elements of the actual implantation process. It will provide practical solutions to a couple that might need to utilise Embryo Glue to meet their conception needs. Basically, the whole process of artificial insemination is based on the need to create a foetus that is a faithful representation of the parents that are undergoing the process (Johansen & Johansen, 2004). There is also the need to be careful in the selection of the genes and the cells that are going to be applied in the process (Anderson, et al., 2004). This implies that there must be the selection of the right embryos to provide the best possible foetus that can withstand the entire gestation process and system. This will help to promote strong reaction and coordination with the substances in the embryonic glue as some schools of thought believe that the embryonic glue is a catalyst that sparks reactions, rather than a unique product in itself. Therefore, there is the need for the selection of the right products to be done in order to achieve the best and most appropriate result. Timing is also an important aspect and feature of the proper and successful utilisation of Embryo Glue. Embryo Glue is a solution that is presented in the loading of the embryos during the transfer process. Therefore, it must be administered within the right and optimal time frame to prevent any damage to the fertilised egg. The eggs are acquired when the patient is induced to super-ovulate for multiple eggs to be produced. These eggs are kept for a period of 9 to 14 days and after that when they are matured, the fertilisation occurs which lasts for another 3-5 days (Kreiner, 2014). In the transfer process, it is advisable to prepare the culture media with Embryo Glue because although the fertilised egg can survive outside the lab conditions for over 1 hour, it is better to just transfer it into the Embryo Glue prepared media immediately to guarantee an adequate volume of proteins for survival. Another aspect of the entire conception process that is important is the washing of the embryo which should be done by the embryologist through various techniques. This involved the gauging of the quantity of cells and other related substances within the local environment of the fertilised egg (Schoolcraft, et al., 2001). This ensures that only the best and the most appropriate quantity of materials surrounds the core of the fertilised egg and this is put in a form whereby it can be applied and presented to the patient. This reduces the risk of foreign materials or excessive detrimental materials going through the conception process. Washing also encompasses the washing of the equipment and clinical tools that will be used in the entire embryonic transfer process. There is the need for the coverslip to be washed in order to eliminate any risks relating to bad results in the microinjection (Jaffe & Terasaki, 2004). This is important because any extra materials in a micro-injection system will lead to major problems and major issues that could alter the genetic composition of the entire fertilised embryo system. It must be recalled that Embryo Glue is seen by some scientists as a supplement rather than the primary ingredient of the process. Therefore, there must be the right local environment in the patient receiving the transplant to achieve the right results. This includes a neutralised pH level that will be around 7.5 to ensure the survival and motion of the foetus (Ly, et al., 2010). Conclusion Embryo Glue is a product on the market that is made up of a primary ingredient, Hyaluronic Acid (HA). This provides a medium and a gel that contains the right environment to promote fertilisation. The main ability of the HA is that it induces adhesion in the CD44 cells of humans that are contained in fertilised eggs that travels into the womb. And the condition created by HA enhances adhesion to the womb and the growth of the foetus by reliance on the nutrients supplied to the walls of the womb. Embryo Glue works best in younger patients and could result in multiple conception. 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