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5 Ultrasound andOvarian Reserve
81

Ovarian Cysts

Ovarian cysts are important to diagnose by ultra­sound in the infertile patient and can affect the ovarian volume. The most common ovarian cysts seen in infertility patients are simple functional cysts, hemorrhagic cysts, endometriomas, and dermoid cysts. Functional cysts are the most common cystic masses seen on ultrasound in the reproductive age group. These cysts tend to stay less than 10 cm and regress after one to two cycles and are either follicular cysts or luteal cysts. If they are small (<3cm) and not hormon­ally active, they do not need to be treated before
Fig. 5.5 (a) Hemorrhagic
cyst. (b) Endometrioma with ground glass appearance. (c) Dermoid cyst 3D with hair. (d) Dermoid cyst 2D seba­ceous material and hair. (e) Malignant ovarian cyst with blood ow into the cyst
ART.However, patients with large simple ovar­ian cysts may have lower response to stimulation, and ovarian cyst aspiration immediately prior to ovarian stimulation has been shown to be bene­cial [42]. A hemorrhagic cyst will show no ow into the cyst, will jiggle with the ultrasound probe, and will resolve spontaneously (Fig.5.5a).
An endometrioma is also a very common nd­ing in the infertile patient and is a sign of the presence of endometriosis in other areas [43, 44]. The typical endometrioma is a unilocular cyst with homogeneous low-level internal echo­genicity (ground glass echogenicity) of the cyst uid (Fig.5.5b). With respect to endometrioma, a
a
b
82
L. A. Stadtmauer et al.
c
d
e
Fig. 5.5 (continued)
5 Ultrasound andOvarian Reserve
83
correct diagnosis is important for infertility with possible need for ART.Transvaginal ultrasonog­raphy is the imaging of choice to differentiate ovarian endometriomas from other adnexal masses [45, 46]. Studies show that an endome­trioma is associated with lower response to ovar­ian stimulation. However, removing the endometrioma can also affect ovarian response and may signicantly diminished ovarian reserve as the base of the ovary is usually burned and the cyst wall is stripped with damage to the follicles [47]. Dermoid cysts can present as solid hyper­echoic heterogeneous masses with a mixed pat­tern of solid and cystic areas (Fig.5.5c, d). They may contain calcications, fat, and hair. Dermoids should be removed prior to IVF if they are caus­ing pain or if there is a question of malignancy. Puncture during oocyte retrieval should be avoided due to high risk of peritonitis.
Although malignancy is rare in women of reproductive age, a suspicious ovarian mass may be seen on an initial infertility scan. Differentiating benign from malignant tumors is done with a combination of morphological evaluation and Doppler ultrasound [48]. The sensitivity and specicity for a combination of US ndings and color Doppler in predicting benign from malig­nant were 0.93 and 0.85in this study. Figure5.5e shows an ovarian mass highly suspicious of a malignancy, a complex mass with papillary pro­jections, and blood ow into the cyst.
Ultrasound andPolycystic Ovary
Polycystic ovary syndrome (PCOS) is one of the most common endocrine disorders impairing female fertility; it has been reported to occur in about 20% of the general female population and in up to 50% of women undergoing IVF therapy [4951]. This topic is covered in detail in another chapter but will be discussed briey here. Ultrasound is one of the criteria for the diagnosis of PCOS. Current data suggest that polycystic ovaries detected by transvaginal ultrasonography may be found in approximately 75% of women with a clinical diagnosis of PCOS [52, 53]. However, it is not a rule that all women with polycystic ovaries will demonstrate the clinical
and biochemical features of PCOS such as oligo­menorrhea and/or hyperandrogenism. Polycystic ovaries per se, even without PCOS, constitute a risk factor for the development of ovarian hyper­stimulation syndrome (OHSS), and the stimula­tion protocol chosen should reect this.
Transvaginal ultrasound is a highly sensitive method for identication of polycystic ovaries. The most commonly used criteria today are those proposed by Dewailly and colleagues with a string of pearls pattern [54, 55] where the antral follicles are arranged peripherally with a dense core of ovarian stroma (Fig. 5.6). This was reafrmed in the Rotterdam 2003 consensus which proposed that the transvaginal threshold for PCOM is based on the presence of an ovarian volume 10cm3 (milliliters) or 12 small folli­cles in a single ovary [56]. The use of 3D ultra­sound and of color and pulsed Doppler ultrasound showing increased ovarian blood ow may fur­ther enable the identication of polycystic ova­ries but is not mandatory for the diagnosis [57]. However, it is important to note that results of studies on ovarian stromal blood ow and vascu­larization in PCOS have demonstrated conict­ing results [58].
With advancement in ultrasound technology and concern that polycystic ovarian morphology (PCOM) may be overdiagnosed using the Rotterdam follicle number per ovary (FNPO) (12 small follicles in a single ovary), the Androgen Excess Society and Polycystic Ovary Syndrome Society (AEPS) 2013 guidelines rec­ommend using the follicle number per ovary ≥25 as the ultrasound threshold for PCOS when using newer technology that has maximal resolution of ovarian follicles (i.e., transducer frequency >8MHz) [59]. The AEPS task force found that studies investigating women from the general population and studies analyzing control and PCOS populations with appropriate statistics suggest setting the criteria for increased FNPO at 25 follicles in women aged 18–35 years but maintaining the criteria for increased ovarian vol­ume at ≥10 cm
3
. They also noted that ovarian volume had less diagnostic potential for PCOM than FNPO.However, if such technology is not available, AEPS recommends using ovarian vol­ume 10cm3 as the ultrasound threshold for rou-
84
Fig. 5.6 Polycystic-
appearing ovary
L. A. Stadtmauer et al.
tine daily practice but not for research inquiries that necessitate the exact and complete character­ization of patients.
Comparisons between transabdominal and transvaginal ultrasound do not nd signicant differences in the detection rate of polycystic ovaries. However, transabdominal sonography is not suitable for acquiring an exact follicle count but may reliably be used to quantify ovarian vol­ume especially in circumstances where it is the only method to evaluate ovarian morphology. The automated 3D US techniques facilitate the exclusion of a false-positive PCOS diagnosis and reect pathophysiological changes in these patients in a more accurate manner. It provides novel means to calculate follicle count and assess for ovarian volume and stromal vascularity. Due to the elevated risk for OHSS, an early detection of PCOS is highly recommended for women undergoing IVF treatment. No other imaging modality, such as magnetic resonance imaging (MRI) for the visualization of the ovaries, is needed for the diagnosis of PCOS.
At present, a few studies have evaluated the use of 3D US in PCOS patients [6069], and a smaller amount has addressed automated 3D US (SonoAVC) [33, 57, 58, 70, 71]. Only one study had postulated a 3D US follicle threshold for PCOM using 3D volume-based software [57]. In this retrospective cohort study, Allemand et al.
analyzed 29 normoandrogenic, ovulatory women with tubal or male factor infertility and 10 PCOS women with chronic anovulation and clinical or biochemical hyperandrogenism. Mean FNPO and the maximal number of follicles in a single sonographic plane (FSSP) were determined by 3D TVUS, and the ovarian volume was deter­mined by 2D TVUS simultaneously. The authors posited a higher threshold of antral follicles (20 or more) for PCOS patients, which is consider­ably higher than that of the Rotterdam criteria. The authors explained this discrepancy by the identication of more follicles by 3D US.Moreover, the authors found the agreement level between the observers was 0.82. However, the weaknesses of this study are the small num­ber of patients, missing direct comparison to 2D US, and low sensitivity (true positive rate) using receiver-operating characteristics (ROCs). Two studies comparing the follicle count between 3D and 2D methodology in polycystic ovaries had conicting results. While Battaglia et al. [64] found no signicant difference in the mean num­ber of follicles between 2D and 3D US calcula­tions in females with polycystic ovaries, Nylander etal. [72] found that 3D US counted more folli­cles than 2D US. The authors also found that ovarian volume measurements by 2D TVUS was
1.48mL smaller than from 3D TVUS.In addi­tion, there is increased stromal blood ow mea-
5 Ultrasound andOvarian Reserve
85
sured by Doppler and decrease in RI into the ovary. This may be part of the enhancement of production of androgens.
According to the AEPS, the literature suggests that 3D ultrasonography holds promise in the assessment of PCOM but states that more studies are needed before AEPS can recommend its rou­tine use.
According to most publications dealing with 3D US in PCOS patients, there is a broad agree­ment about increased ovarian volume of polycys­tic ovaries [64, 66]. According to AEPS, the mean ovarian volume ranges from 10.6 and 16.7mL in female with polycystic ovaries compare to 5.2 to
8.7 mL in healthy female of reproductive age [59]. Pascual etal [63] and Battaglia etal. [64] found strong correlation in ovarian volume calcu­lations between 3D and 2D ultrasonography; however, AEPS stated that the discrepancy in overall ovarian size between the two studies (3D US measurements: 12.6 vs 13.2 P<0.05, respec­tively) was signicant, suggesting technical and interobserver variability. Despite this observa­tion, AEPS did not recommend against using 3D US in determining ovarian volume for the diag­nosis of PCOS.Conclusively, AEPS highly rec­ommends the use of in-house reference normal ovarian volume values but recommends the exist­ing ovarian volume ≥10cm3 criteria determine by either 2D or 3D US if in-house reference nor­mal values are unavailable.
Nevertheless, there is still disagreement regarding ovarian vascularity, stromal changes, and cutoff values for antral follicles. More about PCOS is discussed in Chap. 6.
Antral Follicle Count andSonoAVC
The antral follicle count in 2D and 3D are gener­ally similar although 3D ultrasound is superior for the high antral follicle population. In the inversion mode, follicles will appear white and can be counted easily. Image or volume rotation using the cine-loop facilitates the process (see Fig. 5.2a). Assisted reproductive techniques including invitro fertilization (IVF) and ovula­tion induction require close monitoring of follic-
ular development. Follicular development is commonly assessed with transvaginal ultra­sounds during which each follicle is measured in two or three dimensions. As there are often more than 10–20 follicles, this is very time-consuming. In addition, follicular borders are irregular, and hence, this method has poor reproducibility and is often an inaccurate assessment of follicular volume. A software program called the auto­mated follicular assessment using segmental topology or FAST program was developed to automate the follicular measurements. Therefore, our objective was to determine the accuracy, reproducibility, and efciency of this novel pro­gram. This will be discussed further in another chapter.
One disadvantage of 2D ultrasonography for antral follicle counts involves inter- and intraob­server reproducibility. Of note, only a handful of studies compare the accuracy of ultrasound in regard to AFC and inter-/intraobserver reliability. Scheffer etal. [73] described the value of AFC determined by AVC needed for the improvement of standardization. They compared healthy vol­unteers with proven fertility to patients visiting an infertility clinic. For each patient, 2D or 3D TVUS was conducted for AFC (2–10mm), and interobserver reliability was calculated. Both techniques were adequate when only a few folli­cles were present; however, when higher AFCs occurred, the reproducibility decreased with the 2D technique. In addition to this report, studies by the group of Raine-Fenning [33, 70, 71] dem­onstrated an improvement of interobserver/ intraobserver reliability by the application of 3D methods (by automated systems, such as SonoAVC).
Research has been conducted to evaluate if AFCs vary at different point in the menstrual cycle and if ultrasonographic markers of ovarian reserve differ between the left and right ovary. Interestingly, Deb etal. found that small antral follicles (6.0mm) calculated using 3D US dis­played little intra-cycle variation, while large AFC (>6.0 mm) and ovarian volume revealed signicant intra-cycle variation [12]. Though some studies found there was no signicant dif­ference in AFC between the right and left ovary
86
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[74, 75], most studies, especially those with larger sample sizes and utilizing 3D US, found that AFC was higher in the right ovary than the left [76, 77]. For instance, in a study of 1423 female of reproductive age, Korsholm et al. found that the right ovarian volume was larger than the left, AFC was higher in the right in those with higher AMH, and almost half of those with polycystic ovarian morphology in one ovary, more so in the right ovary than the left [78]. When measured with 3D US, Deb etal. also found that there is a signicant differ­ence in ovarian volume, total AFC of each ovary, and antral follicles >6.0mm between the right and left ovary within an individual [79]. These studies stress the importance of performing a full characterization of both ovaries when scan­ning patients.
The SonoAVC software is especially designed for the automatic detection of multifollicular growth and to overcome the aforementioned problems of follicle observation by 2D US [32,
34, 70, 8086]. This software allows not only the
display of single follicles in 3D but also automat­ically calculates the number and volume of hypoechoic structures in a 3D echo-derived data set; furthermore, it provides estimations regard­ing the absolute dimensions of follicles. The only necessary requirement is adjustment of the ROI box over the entire ovary to include only the information which is needed for calculation and to check during post-processing. Measurements are based on calculation of the largest diameters in three orthogonal planes: the mean follicle diameter, the follicular volume, and the volume­based diameter of the follicle. Thus, the volume is calculated via the voxel count within hypoechoic structures. The true volume can be measured, and SonoAVC calculates the follicle diameter by rendering the follicle as a perfect sphere. This results in the visualization of sono­anatomic details, which could not previously be recognized. A major advantage of this sono­anatomic rendering is that individual structures are color-coded (see Fig.5.4) allowing the exact count, which is needed for the determination of factors such as ovarian reserve. More about SonoAVC is discussed in Chap. 17.
AFC, AMH, andOocyte Cryopreservation
Oocyte or embryo preservation after ovarian stimu­lation, ovarian tissue cryopreservation, invitro mat­uration (IVM) of cumulo-oocyte complexes, and downregulation of ovaries with gonadotropin­releasing hormone analogue during chemotherapy are methods that have been attempted to preserve fertility in the female oncological patient. Though some of these options are controversial or experi­mental, others are more established for fertility preservation. The American Society of Clinical Oncology and the American Society for Reproductive Medicine state that ovarian stimula­tion for embryo or mature oocyte cryopreservation is the most established strategy to result in a future pregnancy [87, 88]. In those with estrogen-sensitive tumors or those who cannot delay cancer therapy, transvaginal retrieval of immature oocytes within IVM of oocytes may be considered. In this method, immature oocytes are retrieved from unstimulated postpubertal ovaries and subsequently matured in the laboratory for mature oocyte or embryo cryo­preservation [88]. In a study by Sonigo etal., the researcher found that 20 antral follicles and 3.7ng/ ml AMH were the thresholds for cryopreservation of at least 10 meiosis II oocytes after IVM, while having ≤19 follicles or 3.0ng/ml AMH was asso- ciated with a risk of cryopreservation of 2 mature oocytes [89]. However, due to limited studies on the safety and efcacy of this approach in the oncologi­cal patient, this technique is still considered investi­gational [88]. More studies need to be done to assess the efcacy and safety of this method and the poten­tial of these oocytes after cryopreservation in patients with malignancies.

Conclusion

AFC is highly predictive for the ovarian reserve reected by the number of oocytes retrieved on stimulation and strongly associated with the serum AMH level. This is helpful for counselling the patient and determining the stimulation pro­tocol. The estimation of the ovarian response by US is simple and noninvasive; thus, it is an advis-
5 Ultrasound andOvarian Reserve
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able procedure. The reliability of this method is increased with 3D instruments. There are several advantages of the new 3D US techniques. Software systems such as SonoAVC allow decreased inter- and intrapersonal variability in the processing, readout, and interpretation of ultrasound scans. While manual post-processing of US scans is required, this can be performed later once the patient is off the table. The ultra­sound denition of PCOS may need modication to incorporate the 3D automated technology. More studies need to be done to evaluate the potential of cryopreservation of ovarian tissue or immature oocytes in the oncological patient.

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